EP4637718A1 - Nanoparticules lipidiques comprenant des phospholipides modifiés par stérol - Google Patents

Nanoparticules lipidiques comprenant des phospholipides modifiés par stérol

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Publication number
EP4637718A1
EP4637718A1 EP23905973.6A EP23905973A EP4637718A1 EP 4637718 A1 EP4637718 A1 EP 4637718A1 EP 23905973 A EP23905973 A EP 23905973A EP 4637718 A1 EP4637718 A1 EP 4637718A1
Authority
EP
European Patent Office
Prior art keywords
lnp
lipid
independently
mol
integer
Prior art date
Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
Pending
Application number
EP23905973.6A
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German (de)
English (en)
Inventor
Dandan LING
Jianxiu XUE
Jerry C. ZHANG
Bo YING
Current Assignee (The listed assignees may be inaccurate. Google has not performed a legal analysis and makes no representation or warranty as to the accuracy of the list.)
Suzhou Abogen Biosciences Co Ltd
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Suzhou Abogen Biosciences Co Ltd
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Publication date
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Publication of EP4637718A1 publication Critical patent/EP4637718A1/fr
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Classifications

    • 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
    • A61K9/10—Dispersions; Emulsions
    • A61K9/127—Synthetic bilayered vehicles, e.g. liposomes or liposomes with cholesterol as the only non-phosphatidyl surfactant
    • A61K9/1271—Non-conventional liposomes, e.g. PEGylated liposomes or liposomes coated or grafted with polymers
    • A61K9/1272—Non-conventional liposomes, e.g. PEGylated liposomes or liposomes coated or grafted with polymers comprising non-phosphatidyl surfactants as bilayer-forming substances, e.g. cationic lipids or non-phosphatidyl liposomes coated or grafted with polymers
    • 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

Definitions

  • the invention relates to lipid nanoparticles comprising phospholipids that contain a sterol moiety.
  • LNPs Lipid nanoparticles
  • mRNA vaccines One important application of nucleic acid-loaded LNPs are mRNA vaccines.
  • the present application provides phospholipids containing a sterol moiety that can be used to construct lipid nanoparticles.
  • the lipid nanoparticles are useful for delivering nucleic acids, e.g. mRNA, to one or more cells.
  • the application provides a method for expressing protein in a cell by delivering nucleic acids to the cell via the lipid nanoparticle comprising a phospholipid that contains a sterol moiety.
  • lipid nanoparticles comprising
  • the phospholipid has a structure selected from:
  • the phospholipid has the structure:
  • the phospholipid has the structure:
  • the LNP has a molar ratio of the ionizable lipid to the phospholipid from 20 ⁇ 1 to 2 ⁇ 1. In some embodiments, the molar ratio of ionizable lipid to phospholipid is from 15 ⁇ 1 to 5 ⁇ 1. In some embodiments, the i onizable lipid comprises from 40 to 80 mol%of a total amount of lipids in the LNP. In some embodiments, the ionizable lipid comprises from 50 to 70 mol%of the total amount of lipids in the LNP. In some embodiments, the ionizable lipid is a cationic lipid.
  • the ionizable lipid is a compound according to any one of the formula selected from 01-I, 01-II, 02-I, 02-II, 03-I, 03-II-A, 03-II-B, 03-II-C, 03-II-D, 04-I, 04-III, 04-IV, 05-I, 06-I, and sub-formulas thereof, or wherein the ionizable lipid is a cationic lipid selected from the compounds listed in any one of Tables 1 to 5.
  • the polymer conjugated lipid comprises from 0.5 to 5 mol%of the total amount of lipids in the LNP. In some embodiments, the polymer conjugated lipid comprises from 1 to 2 mol%of the total amount of lipids in the LNP. In some embodiments, the polymer conjugated lipid comprises 1.5 mol%of the total amount of lipids in the LNP. In some embodiments, the LNP has a molar ratio of the polymer conjugated lipid to the phospholipid of from 1 ⁇ 2 to 1 ⁇ 20. In some embodiments, the LNP has a molar ratio of the polymer conjugated lipid to the phospholipid of from 1 ⁇ 3 to 1 ⁇ 18.
  • the LNP has a molar ratio of the polymer conjugated lipid to the phospholipid from 1 ⁇ 5 to 1 ⁇ 10.
  • the polymer conjugated lipid is a PEGylated lipid.
  • the polymer conjugated lipid is a PEGylated lipid with the structure:
  • R 12 and R 13 are each independently a straight or branched, alkyl or alkenyl chain containing from 10 to 30 carbon atoms, wherein the alkyl chain is optionally interrupted by one or more ester bonds;
  • w is an integer ranging from 30 to 60.
  • the polymer conjugated lipid is a PEGylated lipid with the structure:
  • w is an integer ranging from 30 to 60.
  • w is an integer ranging from 45 to 55.
  • w is about 49.
  • the polymer conjugated lipid is DMG-PEG or DMPE-PEG.
  • the LNP further comprises a lipid stabilizer.
  • the LNP has a molar ratio of the lipid stabilizer to the phospholipid of from 10 ⁇ 1 to 1 ⁇ 4. In some embodiments, the molar ratio of the lipid stabilizer to the phospholipid is from 5 ⁇ 1 to 1 ⁇ 3. In some embodiments, the LNP has a molar ratio of the lipid stabilizer to the phospholipid of from 10 ⁇ 1 to 1 ⁇ 2. In some embodiments, the molar ratio of the lipid stabilizer to the phospholipid is from 5 ⁇ 1 to 1 ⁇ 1. In some embodiments, the molar ratio of the lipid stabilizer to the phospholipid is from 4 ⁇ 1 to 3 ⁇ 1.
  • the lipid stabilizer comprises from 5 to 50 mol%of the total amount of lipids in the LNP. In some embodiments, the lipid stabilizer comprises from 8 to 40 mol%of the total amount of lipids in the LNP. In some embodiments, the lipid stabilizer comprises from 10 to 30 mol%of the total amount of lipids in the LNP.
  • the phospholipid comprises from 1 to 30 mol%of the total amount of lipids in the LNP. In some embodiments, the phospholipid comprises from 2 to 25 mol%of the total amount of lipids in the LNP. In some embodiments, the phospholipid comprises from 3 to 20 mol%of the total amount of lipids in the LNP. In some embodiments, the phospholipid comprises from 5 to 15 mol%of the total amount of lipids in the LNP. In some embodiments, the phospholipid comprises about 10 mol%of the total amount of lipids in the LNP.
  • the LNP has a size of from 20 nm to 300 nm, as determined using dynamic light scattering. In some embodiments, the LNP has a size of from 50 nm to 150 nm, as determined using dynamic light scattering. In some embodiments, the size is from 60 nm to 140 nm. In some embodiments, the size is from 80 nm to 100 nm. In some embodiments, the size is from 85 nm to 95 nm.
  • the LNP encapsulates mRNA.
  • LNPs comprising a phospholipid, wherein the phospholipid has a structure:
  • the phospholipid has the structure:
  • the phospholipid has the structure:
  • the LNP further comprises an ionizable lipid.
  • the LNP has a molar ratio of the ionizable lipid to the phospholipid from 20 ⁇ 1 to 2 ⁇ 1. In some embodiments, the molar ratio of ionizable lipid to phospholipid is from 15 ⁇ 1 to 5 ⁇ 1. In some embodiments, the i onizable lipid comprises from 40 to 80 mol%of a total amount of lipids in the LNP. In some embodiments, the ionizable lipid comprises from 50 to 70 mol%of the total amount of lipids in the LNP.
  • the ionizable lipid is a compound according to any one of the formula selected from 01-I, 01-II, 02-I, 02-II, 03-I, 03-II-A, 03-II-B, 03-II-C, 03-II-D, 04-I, 04-III, 04-IV, 05-I, 06-I, and sub-formulas thereof, or wherein the ionizable lipid is a cationic lipid selected from the compounds listed in any one of Tables 1 to 5. In some embodiments, the ionizable lipid is a cationic lipid.
  • the LNP further comprises a polymer conjugated lipid.
  • the polymer conjugated lipid comprises from 1 to 2 mol%of a total amount of lipids in the LNP. In some embodiments, the polymer conjugated lipid comprises 1.5%of the total amount of lipids in the LNP.
  • the LNP has a molar ratio of the polymer conjugated lipid to the phospholipid from 1 ⁇ 5 to 1 ⁇ 10. In some embodiments, the polymer conjugated lipid is a PEGylated lipid.
  • the polymer conjugated lipid is a PEGylated lipid with the structure:
  • R 12 and R 13 are each independently a straight or branched, alkyl or alkenyl chain containing from 10 to 30 carbon atoms, wherein the alkyl chain is optionally interrupted by one or more ester bonds;
  • w is an integer ranging from 30 to 60.
  • the polymer conjugated lipid is a PEGylated lipid with the structure:
  • w is an integer ranging from 30 to 60.
  • w is an integer ranging from 45 to 55. In some embodiments, w is about 49.
  • the polymer conjugated lipid is DMG-PEG or DMPE-PEG.
  • the LNP further comprises a lipid stabilizer.
  • the LNP has a molar ratio of the lipid stabilizer to the phospholipid from 10 ⁇ 1 to 1 ⁇ 4. In some embodiments, the molar ratio of the lipid stabilizer to the phospholipid is from 5 ⁇ 1 to 1 ⁇ 3. In some embodiments, the molar ratio of the lipid stabilizer to the phospholipid is from 4 ⁇ 1 to 3 ⁇ 1. In some embodiments, the lipid stabilizer comprises from 5 to 50 mol%of a total amount of lipids in the LNP. In some embodiments, the lipid stabilizer comprises from 8 to 40 mol%of the total amount of lipids in the LNP. In some embodiments, the lipid stabilizer comprises from 10 to 30 mol%of the total amount of lipids in the LNP.
  • the phospholipid comprises from 1 to 30 mol%of a total amount of lipids in the LNP. In some embodiments, the phospholipid comprises from 2 to 25 mol%of the total amount of lipids in the LNP. In some embodiments, the phospholipid comprises from 3 to 20 mol%of the total amount of lipids in the LNP. In some embodiments, the phospholipid comprises from 5 to 15 mol%of the total amount of lipids in the LNP. In some embodiments, the phospholipid comprises about 10 mol%of the total amount of lipids in the LNP. In some embodiments, the LNP has a size of from 20 nm to 300 nm, as determined using dynamic light scattering.
  • the LNP has a size of from 50 nm to 150 nm, as determined using dynamic light scattering. In some embodiments, the size is from 60 nm to 140 nm. In some embodiments, the size is from 80 nm to 100 nm. In some embodiments, the size is from 85 nm to 95 nm.
  • the LNP encapsulates mRNA.
  • compositions of the lipid nanoparticles (LNPs) described herein are also provided herein.
  • LNPs lipid nanoparticles
  • at least 80%of the LNPs encapsulate mRNA.
  • at least 85%of the LNPs encapsulate mRNA.
  • the cell is a mammalian cell.
  • the LNP or composition thereof is administered systemically.
  • the subject is a mammal. In some embodiments, the subject is a human.
  • FIG. 1 shows expression levels of hEPO in LNP formulations containing PChemsPC or OChemsPC with a 60 ⁇ 10 ⁇ 28.5 ⁇ 1.5 molar ratio of compound 01-1/OChemsPC or PChemsPC/Chol/DMG-PEG in Example 15.
  • FIG. 2 shows expression levels of hEPO in LNP formulations containing DSPC or PChemsPC at various molar percentages in Example 16.
  • FIG. 3 shows expression levels of hEPO in LNP formulations containing PChemsPC at various molar ratios of PChemsPC/Chol in Example 17.
  • FIG. 4 shows the luminescence levels measured from harvest liver tissues in Example 19.
  • Fig. 5 shows the percentage of luminescence intensity in different tissues in Example 19.
  • FIG. 6 shows hEPO expression fold change of LNPs with or without a steroid containing phospholipid in Example 20.
  • FIG. 7 shows serum cytokines levels boosted by compound 01-1 LNP in Example 21.
  • FIG. 8 shows serum cytokines levels boosted by Lipid 5 LNP in Example 21.
  • FIG. 9 shows serum cytokines levels boosted by SM-102 LNP in Example 21.
  • FIG. 11 shows serum cytokines levels boosted by Compound 03-135 LNP in Example 21.
  • FIG. 12 shows serum cytokines levels boosted by Compound 01-1 saRNA-LNP in Example 22.
  • FIG. 13 shows serum cytokines levels boosted by Compound 03-135 saRNA-LNP in Example 22.
  • FIG. 14 shows CD3-CD19 antibody levels of LNPs with or without a steroid containing phospholipid in Example 23.
  • LNPs comprising a phospholipid containing a sterol moiety.
  • the LNPs can be loaded with mRNA, such as in mRNA vaccine technology.
  • Sterol-modified phospholipids stabilize bilayers but do not exchange between membranes as freely as cholesterol.
  • the mRNA-loaded LNPs demonstrate an ability to increase protein expression in target cells compared to mRNA-loaded LNPs with more conventional phospholipids. Increasing protein expression helps increase the effectiveness and efficiency of mRNA-based treatments and therapies.
  • pharmaceutically acceptable or “pharmacologically compatible” is meant a material that is not biologically or otherwise undesirable, e.g., the material may be incorporated into a pharmaceutical composition administered to a patient without causing any significant undesirable biological effects or interacting in a deleterious manner with any of the other components of the composition in which it is contained.
  • Pharmaceutically acceptable carriers or excipients have preferably met the required standards of toxicological and manufacturing testing and/or are included on the Inactive Ingredient Guide prepared by the U.S. Food and Drug administration.
  • a pharmaceutically acceptable carrier refers to a pharmaceutically acceptable substrate, composition or vehicle used in the process of drug delivery, which may have one or more ingredients including, but not limited to, excipient (s) , binder (s) , diluent (s) , solvent (s) , filler (s) , and/or stabilizer (s) .
  • lipid refers to a group of compounds including, without limitation, fats, sterols, waxes, fat-soluble vitamins, monoglycerides, diglycerides, sphingolipids, and phospholipids.
  • phospholipids, ionizable lipids, polymer conjugated lipids, and lipid stabilizers are considered lipids.
  • the term “ionizable lipid” refers to a lipid that has a non-zero net electric charge at physiological pH.
  • the term is inclusive with respect to cationic lipids, including lipids that have a partial positive charge at physiological pH.
  • the term is also inclusive with respect to mixtures of ionizable lipids, which can contain two or more ionizable lipids.
  • ionizable lipid it is likewise contemplated with a “cationic lipid, ” as though all embodiments were specifically and individually listed with both ionizable and cationic lipids.
  • polymer conjugated lipid refers to a lipid comprising a polymer moiety.
  • the term is inclusive with respect to PEGylated lipids, including PEGylated phosphatidylethanolamines, PEGylated phosphatidic acids, PEGylated ceramides, PEGylated dialkylamines, PEGylated diacylglycerols, and PEGylated dialkylglycerols.
  • the term is also inclusive with respect to mixture of polymer conjugated lipids, which may contain two or more polymer conjugated lipids.
  • lipid stabilizer refers to a component of the lipid nanoparticle that thought to help stabilize the LNP structure. Without being bound by theory, it is believed that the lipid stabilizer component of LNPs helps favor the liquid-ordered phase of the lipid membrane in LNPs. See, for example, section 3.3.1 of Albertsen, H.C.; et al., “The role of lipid components in lipid nanoparticles for vaccines and gene therapy. ” Adv Drug Deliv Rev. 2022 Sep; 188: 114416. Compounds that can serve as lipid stabilizers include sterols, corticosteroids, vitamins, and other compounds comprising a steroid core.
  • alkyl refers to a chain of carbon atoms wherein all bonds between the carbon atoms in the alkyl group are single bonds.
  • the term is inclusive with respect to straight and branched chains (e.g., the term includes both n-propyl and isopropyl groups) .
  • C x -C y alkyl refers to an alkyl with at least x carbon atoms and no more than y carbon atoms in the alkyl chain.
  • C 1 -C 3 alkyl includes, without limitation, methyl, ethyl, n-propyl, and isopropyl.
  • alkylene refers to an alkyl chain that connects in at least two locations to other chemical groups.
  • C x -C y alkylene refers to an alkylene with at least x carbon atoms and no more than y carbon atoms in the alkylene chain.
  • C 1 -C 3 alkylene includes, without limitation, methylene, ethylene, n-propylene, and iso-propylene.
  • alkenyl refers to a chain of carbon atoms with at least one double bond between two carbon atoms in the chain.
  • the term is inclusive with respect to straight and branched chains (e.g., the term includes both 1 -propenyl and iso-propenyl groups) .
  • alkenylene refers to an alkenyl chain that connects in at least two locations to other chemical groups.
  • C x -C y alkenylene refers to an alkenylene with at least x carbon atoms and no more than y carbon atoms.
  • alkynyl refers to a chain of carbon atoms with at least one triple bond between two carbon atoms in the chain.
  • the term is inclusive with respect to straight and branched chains (e.g., the term includes both 1-propynyl and iso-propynyl groups) .
  • C x -C y alkynyl refers to an alkynyl with at least x carbon atoms and no more than y carbon atoms in the alkynyl chain.
  • cycloalkyl refers to a cyclic group of carbon atoms wherein all the bonds between the carbon atoms are single bonds.
  • C x -C y cycloalkyl refers to a cycloalkyl with at least x carbon atoms and no more than y carbon atoms.
  • C 6 -C 10 cycloalkyl includes, without limitation, cyclohexyl and cyclo-octyl.
  • cycloalkylene has the same meaning as cycloalkyl, except that the cycloalkylene connects to at least two other chemical groups.
  • cycloalkenyl refers to a cyclic group of carbon atoms where at least one bond between two carbon atoms in the cycloalkenyl group is a double bond.
  • C x -C y cycloalkenyl refers to a cycloalkenyl with at least x carbon atoms and no more than y carbon atoms.
  • C x -C y aryl refers to an aryl with at least x carbon atoms and no more than y carbon atoms.
  • C 6 -C 10 aryl includes, without limitation, phenyl and naphthyl.
  • arylene has the same meaning as aryl, except that the arylene connects to at least two other chemical groups.
  • heterocycloalkyl refers to a cyclic group of atoms wherein all the bonds between the atoms in the ring are single bonds.
  • C x -C y heterocycloalkyl refers to a heterocycloalkyl with at least x atoms and no more than y atoms.
  • C 5 -C 6 heterocycloalkyl includes, without limitation, pyrrolidinyl and 1, 4-dioxanyl.
  • heterocycloalkylene has the same meaning as heterocycloalkyl, except that the heterocycloalkylene connects to at least two other chemical groups.
  • x-to y-membered heteroaryl refers to a cyclic group of atoms with at least x atoms and no more than y atoms.
  • 5-or 6-membered heteroaryl includes, without limitation, pyridinyl and furanyl.
  • the term “carbocycle” refers to a cycloalkyl or an aryl group. Likewise, the term “heterocycle” refers to a heterocycloalkyl or a heteroaryl group.
  • Possible atoms that make up the ring in heterocycloalkyl and heteroaryl groups, as well as derivatives thereof, include, without limitation, carbon, nitrogen, oxygen, and sulfur.
  • the term “optionally substituted” means the indicated group may be substituted or unsubstituted.
  • substituted refers to another chemical moiety that decorates the indicated group by replacement of one H atom.
  • ethanol is an example of ethane substituted with OH.
  • a group that is optionally substituted is optionally substituted by chloro, fluoro, bromo, C 1 -C 6 alkyl, C 2 -C 6 alkenyl, C 2 -C 6 alkynyl, C 3 -C 7 cycloalkyl, C 6 -C 10 aryl, or 5-or 6-membered heteroaryl.
  • the terms “individual, ” “subject, ” and “patient” are used interchangeably herein to describe a mammal, including humans.
  • the individual is in need of treatment, for example, the individual may have been diagnosed with, or is suspected of having, a cancer.
  • references to "about” a value or parameter herein includes (and describes) variations that are directed to that value or parameter per se.
  • description referring to "about X” includes description of "X” .
  • the term “about” a value or parameter means a range within 20%, in either direction, of the value or parameter recited.
  • the mole percentages of lipids and lipid stabilizers in lipid nanoparticles are calculated based on the total mole number of components in the lipid nanoparticles.
  • LNPs Lipid Nanoparticles
  • the lipid nanoparticles (LNPs) herein comprise a phospholipid containing a sterol moiety and optionally one or more of an ionizable lipid, a polymer conjugated lipid, and a lipid stabilizer.
  • the LNPs comprise a phospholipid containing a sterol moiety, an ionizable lipid, and a polymer conjugated lipid.
  • the LNPs comprise a phospholipid containing a sterol moiety, a polymer conjugated lipid, and a lipid stabilizer.
  • the LNPs comprise a phospholipid containing a sterol moiety, an ionizable lipid, and a lipid stabilizer.
  • the LNPs comprise a phospholipid containing a sterol moiety, an ionizable lipid, a polymer conjugated lipid, and a lipid stabilizer.
  • the phospholipid comprises from 1 to 30 mol%of the total lipids in the LNP. In some embodiments, the phospholipid comprises from 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, or 30 mol%of the total lipids in the LNP. In some embodiments, the phospholipid comprises from 2 to 25 mol%of the total lipids in the LNP. In some embodiments, the phospholipid comprises from 3 to 20 mol%of the total lipids in the LNP. In some embodiments, the phospholipid comprises from 5 to 10, 15, 20, 25, or 30 mol%of the total lipids in the LNP.
  • the phospholipid comprises from 10 to 15, 20, 25, or 30 mol%of the total lipids in the LNP. In some embodiments, the phospholipid comprises from 15 to 20, 25, or 30 mol%of the total lipids in the LNP. In some embodiments, the phospholipid comprises from 20 to 25 or 30 mol%of the total lipids in the LNP. In some embodiments, the phospholipid comprises from 25 to 30 mol%of the total lipids in the LNP.
  • the ionizable lipid comprises from 40 to 80 mol%of the total lipids in the LNP. In some embodiments, the ionizable lipid comprises from 40 to 50, 60, 70, or 80 mol%of the total lipids in the LNP. In some embodiments, the ionizable lipid comprises from 45 to 50, 60, 70, 75, or 80 mol%of the total lipids in the LNP. In some embodiments, the ionizable lipid comprises from 50 to 60, 70, or 80 mol%of the total lipids in the LNP. In some embodiments, the ionizable lipid comprises from 60 to 65, 70 or 80 mol%of the total lipids in the LNP.
  • the ionizable lipid comprises from 70 to 80 mol%of the total lipids in the LNP. In some embodiments, the ionizable lipids comprise about 40, 50, 60, 70, or 80 mol%of the total lipids in the LNP. In some embodiments, the ionizable lipids comprise about 50, 60, or 70 mol%of the total lipids in the LNP.
  • the LNP has a molar ratio of the ionizable lipid to the phospholipid of from 20 ⁇ 1 to 2 ⁇ 1. In some embodiments, the molar ratio is from 20 ⁇ 1 to 15 ⁇ 1, 10 ⁇ 1, 5 ⁇ 1, or 2 ⁇ 1. In some embodiments, the molar ratio is from 18 ⁇ 1 to 2.5 ⁇ 1. In some embodiments, the molar ratio is from 16 ⁇ 1 to 4 ⁇ 1. In some embodiments, the molar ratio is from 15 ⁇ 1 to 10 ⁇ 1, 5 ⁇ 1, or 2 ⁇ 1. In some embodiments, the molar ratio is from 10 ⁇ 1 to 5 ⁇ 1 or 2 ⁇ 1. In some embodiments, the ratio is from 5 ⁇ 1 to 2 ⁇ 1. In some embodiments, the ratio is from 15 ⁇ 1 to 5 ⁇ 1.
  • the polymer conjugated lipid has a molar ratio of 0.5 to 5 mol%of the total lipids in the LNP. In some embodiments, the polymer conjugated lipid has a molar ratio of 1 to 2 mol%of the total lipids in the LNP. In some embodiments, the polymer conjugated lipid has a molar ratio of 1.5 mol%of the total lipids in the LNP.
  • the molar ratio of the polymer conjugated lipid to the phospholipid from 1 ⁇ 2 to 1 ⁇ 20. In some embodiments, the molar ratio of the polymer conjugated lipid to the phospholipid from 1 ⁇ 3 to 1 ⁇ 18. In some embodiments, the molar ratio of the polymer conjugated lipid to the phospholipid from 1 ⁇ 5 to 1 ⁇ 10.
  • the lipid stabilizer comprises from 5 to 50 mol%of the total lipids in the LNP. In some embodiments, the lipid stabilizer comprises from 5 to 10, 20, 30, 40, or 50 mol%of the total lipids in the LNP. In some embodiments, the lipid stabilizer comprises from 8 to 20, 30, 40, or 50 mol%of the total lipids in the LNP. In some embodiments, the lipid stabilizer comprises from 10 to 20, 30, 40, or 50 mol%of the total lipids in the LNP. In some embodiments, the lipid stabilizer comprises from 20 to 30, 40, or 50 mol%of the total lipids in the LNP.
  • the lipid stabilizer comprises from 30 to 40 or 50 mol%of the total lipids in the LNP. In some embodiments, the lipid stabilizer comprises from 40 to 50 mol%of the total lipids in the LNP. In some embodiments, the lipid stabilizer comprises about 5, 10, 20, 30, 40, or 50 mol%of the total lipids in the LNP.
  • any of the molar percentages or molar ratios described above for the sterol-containing phospholipid, the ionizable lipid, the polymer conjugated lipid, and the lipid stabilizer may be combined with each other in any embodiment describing the lipid composition of the LNPs described herein, such that every combination is contemplated as though each and every combination were specifically and individually disclosed.
  • the phospholipid comprises from 1 to 30 mol%, the ionizable lipid comprises from 40 to 80 mol%, the polymer conjugated lipid comprises from 1 to 2 mol%, and the lipid stabilizer comprises from 5 to 50 mol%of the total lipids in the LNP. In some embodiments, the phospholipid comprises from 5 to 25 mol%, the ionizable lipid comprises from 45 to 75 mol%, the polymer conjugated lipid comprises from 1 to 2 mol%, and the lipid stabilizer comprises from 20 to 40%of the total lipids in the LNP.
  • the phospholipid comprises from 5 to 15 mol%, the ionizable lipid comprises from 40 to 60 mol%, the polymer conjugated lipid comprises from 1 to 2 mol%, and the lipid stabilizer comprises from 20 to 40 mol%of the total lipids in the LNP. In some embodiments, the phospholipid comprises about 10 mol%, the ionizable lipid comprises about 50 mol%, the polymer conjugated lipid comprises about 38.5 mol%, and the lipid stabilizer comprises about 1.5 mol%of the total lipids in the LNP.
  • the LNP has a size from 20 to 40, 50, 60, 70, 80, 90, 100, 110, 120, 130, 140, 150, 200, 250, or 300 nm. In some embodiments, the size is from 40 to 50, 60, 70, 80, 90, 100, 110, 120, 130, 140, 150, 200, 250, or 300 nm. In some embodiments, the size is from 50 to 60, 70, 80, 90, 100, 110, 120, 130, 140, 150, 200, 250, or 300 nm. In some embodiments, the size is from 60 to 70, 80, 90, 100, 110, 120, 130, 140, 150, 200, 250, or 300 nm.
  • the size is from 70 to 80, 90, 100, 110, 120, 130, 140, 150, 200, 250, or 300 nm. In some embodiments, the size is from 80 to 90, 100, 110, 120, 130, 140, 150, 200, 250, or 300 nm. In some embodiments, the size is from 90 to 100, 110, 120, 130, 140, 150, 200, 250, or 300 nm. In some embodiments, the size is from 100 to 110, 120, 130, 140, 150, 200, 250, or 300 nm. In some embodiments, the size is from 110 to 120, 130, 140, 150, 200, 250, or 300 nm. In some embodiments, the size is from 120 to 130, 140, 150, 200, 250, or 300 nm. In some embodiments, the size is from 120 to 130, 140, 150, 200, 250, or 300 nm. In some embodiments, the size is from 120 to 130, 140, 150, 200, 250, or 300 nm.
  • the size is from 130 to 140, 150, 200, 250, or 300 nm. In some embodiments, the size is from 140 to 150, 200, 250, or 300 nm. In some embodiments, the size is from 150 to 200, 250, or 300 nm. In some embodiments, the size is from 200 to 300 nm. In some embodiments, the size is from 60 to 150 nm. In some embodiments, the size is from 65 to 90 nm. In some embodiments, the size is from 70 to 80 nm. In some embodiments, the size is about 60, 65, 70, 75, 80, 85, 90, 95, 100, 105, 110, or 115 nm. In some embodiments, the size is about 85 or about 90 nm. In some embodiments, the size is about 85 nm. In some embodiments, the size is about 90 nm. In some embodiments, the size is about 90 nm. In some embodiments, the size is about 90 nm. In some embodiments, the size is about 90 nm.
  • the LNPs herein comprise a phospholipid containing a sterol moiety.
  • the phospholipid containing a sterol moiety is any phospholipid that incorporates a sterol moiety into the lipid structure.
  • the sterol moiety is incorporated in place of one or more alkyl chains on the phospholipid.
  • the sterol moiety is incorporated into one alkyl chain of the phospholipid.
  • the sterol moiety is connected through the O atom of the sterol (e.g. converting the sterol moiety into the -O-atom of an ester connection to the remainder of the phospholipid) .
  • the sterol moiety is cholesterol.
  • the sterol moiety is a cholesterol moiety connected through O atom of the sterol (e.g. by converting the sterol O atom of cholesterol into the -O-atom of an ester connection to the remainder of the phospholipid) .
  • the phospholipid has a structure of selected from:
  • the LNPs comprise an ionizable lipid.
  • the ionizable lipid is a cationic lipid.
  • the cationic lipid is a cationic lipid described in International Patent Publication No. WO 2021/204175, the entirety of which is incorporated herein by reference.
  • the cationic lipid is a compound of Formula (01-I) :
  • G 1 and G 2 are each independently a bond, C 2 -C 12 alkylene, or C 2 -C 12 alkenylene, wherein one or more -CH 2 -in the alkylene or alkenylene is optionally replaced by -O-;
  • R 1 and R 2 are each independently C 6 -C 32 alkyl or C 6 -C 32 alkenyl
  • R a , R b , R d , and R e are each independently H, C 1 -C 24 alkyl, or C 2 -C 24 alkenyl;
  • R c and R f are each independently C 1 -C 32 alkyl or C 2 -C 32 alkenyl
  • G 3 is C 2 -C 24 alkylene, C 2 -C 24 alkenylene, C 3 -C 8 cycloalkylene, or C 3 -C 8 cycloalkenylene;
  • R 3 is -N (R 4 ) R 5 ;
  • R 4 is C 3 -C 8 cycloalkyl, C 3 -C 8 cycloalkenyl, 4-to 8-membered heterocyclyl, or C 6 -C 10 aryl; or R 4 , G 3 or part of G 3 , together with the nitrogen to which they are attached form a cyclic moiety;
  • R 5 is C 1 -C 12 alkyl or C 3 -C 8 cycloalkyl; or R 4 , R 5 , together with the nitrogen to which they are attached form a cyclic moiety;
  • x 0, 1, or 2;
  • alkyl, alkenyl, cycloalkyl, cycloalkenyl, heterocyclyl, aryl, alkylene, alkenylene, cycloalkylene, cycloalkenylene, arylene, heteroarylene, and cyclic moiety is independently optionally substituted.
  • the cationic lipid is a compound of Formula (01-II) :
  • G 1 and G 2 are each independently a bond, C 2 -C 12 alkylene, or C 2 -C 12 alkenylene, wherein one or more -CH 2 -in the alkylene or alkenylene is optionally replaced by -O-;
  • R 1 and R 2 are each independently C 6 -C 32 alkyl or C 6 -C 32 alkenyl
  • R a , R b , R d , and R e are each independently H, C 1 -C 24 alkyl, or C 2 -C 24 alkenyl;
  • R c and R f are each independently C 1 -C 32 alkyl or C 2 -C 32 alkenyl
  • G 4 is a bond, C 1 -C 23 alkylene, C 2 -C 23 alkenylene, C 3 -C 8 cycloalkylene, or C 3 -C 8 cycloalkenylene;
  • R 3 is -N (R 4 ) R 5 ;
  • R 4 is C 1 -C 12 alkyl, C 3 -C 8 cycloalkyl, C 3 -C 8 cycloalkenyl, 4-to 8-membered heterocyclyl, or C 6 -C 10 aryl; or R 4 , G 3 or part of G 3 , together with the nitrogen to which they are attached form a cyclic moiety;
  • R 5 is C 1 -C 12 alkyl or C 3 -C 8 cycloalkyl; or R 4 and R 5 , together with the nitrogen to which they are attached form a cyclic moiety;
  • x 0, 1, or 2;
  • alkyl, alkenyl, cycloalkyl, cycloalkenyl, heterocyclyl, aryl, alkylene, alkenylene, cycloalkylene, cycloalkenylene, arylene, heteroarylene, and cyclic moiety is independently optionally substituted.
  • the cationic lipid is a compound of Formula (01-I-B) , (01-I-B’) , (01-I-B”) , (01-I-C) , (01-I-D) , or (01-I-E) :
  • G 1 and G 2 are each independently C 3 -C 7 alkylene. In some embodiments, G 1 and G 2 are each independently C 5 alkylene. In some embodiments, G 3 is C 2 -C 4 alkylene. In some embodiments, G 3 is C 2 alkylene. In some embodiments, G 3 is C 4 alkylene.
  • R 3 has one of the following structures:
  • R 1 , R 2 , R c , and R f are each independently branched C 6 -C 32 alkyl or branched C 6 -C 32 alkenyl. In some embodiments, R 1 , R 2 , R c , and R f are each independently branched C 6 -C 24 alkyl or branched C 6 -C 24 alkenyl. In some embodiments, R 1 , R 2 , R c , and R f are each independently -R 7 -CH (R 8 ) (R 9 ) , wherein R 7 is C 0 -C 5 alkylene, and R 8 and R 9 are independently C 2 -C 10 alkyl.
  • R 1 , R 2 , R c , and R f are each independently -R 7 -CH (R 8 ) (R 9 ) , wherein R 7 is C 0 -C 1 alkylene, and R 8 and R 9 are independently C 4 -C 8 alkyl.
  • the cationic lipid is a compound in Table 1, or a pharmaceutically acceptable salt, prodrug, or stereoisomer thereof.
  • the cationic lipid is a cationic lipid described in International Patent Application No. PCT/CN2022/072694, the entirety of which is incorporated herein by reference. In some embodiments, the cationic lipid is a compound of Formula (02-I) :
  • R 1 and R 2 are each independently C 6 -C 24 alkyl or C 6 -C 24 alkenyl
  • R a , R b , R d , and R e are each independently H, C 1 -C 24 alkyl, or C 2 -C 24 alkenyl;
  • R c and R f are each independently C 1 -C 24 alkyl or C 2 -C 24 alkenyl
  • G 3 is C 2 -C 12 alkylene or C 2 -C 12 alkenylene, wherein part or all of alkylene or alkenylene is optionally replaced by a C 3 -C 8 cycloalkylene or C 3 -C 8 cycloalkenylene;
  • R 3 is -N (R 4 ) R 5 , -OR 6 , or-SR 6 ;
  • R 4 is C 1 -C 12 alkyl, C 2 -C 12 alkenyl, C 3 -C 8 cycloalkyl, C 3 -C 8 cycloalkenyl, C 6 -C 10 aryl, or 4-to 8-membered heterocycloalkyl;
  • R 5 is H, C 1 -C 12 alkyl, C 3 -C 8 cycloalkyl, C 3 -C 8 cycloalkenyl, C 6 -C 10 aryl, or 4-to 8-membered heterocycloalkyl;
  • R 6 is hydrogen, C 1 -C 12 alkyl, C 3 -C 8 cycloalkyl, C 3 -C 8 cycloalkenyl, or C 6 -C 10 aryl;
  • x 0, 1, or 2;
  • each alkyl, alkenyl, cycloalkyl, cycloalkenyl, heterocycloalkyl, aryl, alkylene, alkenylene, cycloalkylene, and cycloalkenylene is independently optionally substituted.
  • the cationic lipid is a compound of Formula (02-II) :
  • R 1 and R 2 are each independently C 6 -C 24 alkyl or C 6 -C 24 alkenyl
  • R a , R b , R d , and R e are each independently H, C 1 -C 24 alkyl, or C 2 -C 24 alkenyl;
  • R c and R f are each independently C 1 -C 24 alkyl or C 2 -C 24 alkenyl
  • G 3 is C 2 -C 12 alkylene or C 2 -C 12 alkenylene, wherein part or all of alkylene or alkenylene is optionally replaced by a C 3 -C 8 cycloalkylene or C 3 -C 8 cycloalkenylene;
  • R 3 is -N (R 4 ) R 5 , -OR 6 , or-SR 6 ;
  • R 4 is C 1 -C 12 alkyl, C 2 -C 12 alkenyl, C 3 -C 8 cycloalkyl, C 3 -C 8 cycloalkenyl, C 6 -C 10 aryl, or 4-to 8-membered heterocycloalkyl;
  • R 5 is H, C 1 -C 12 alkyl, C 3 -C 8 cycloalkyl, C 3 -C 8 cycloalkenyl, C 6 -C 10 aryl, or 4-to 8-membered heterocycloalkyl;
  • R 6 is hydrogen, C 1 -C 12 alkyl, C 3 -C 8 cycloalkyl, C 3 -C 8 cycloalkenyl, or C 6 -C 10 aryl;
  • x 0, 1, or 2;
  • each alkyl, alkenyl, cycloalkyl, cycloalkenyl, heterocycloalkyl, aryl, alkylene, alkenylene, cycloalkylene, and cycloalkenylene is independently optionally substituted.
  • the compound is a compound of Formula (02-V-A) , (02-V-B) , (02-V-C) , (02-V-D) , (02-V-E) , (02-V-F) :
  • z is an integer from 2 to 12
  • x0 is an integer from 1 to 11;
  • y0 is an integer from 1 to 11;
  • x1 is an integer from 0 to 9;
  • y1 is an integer from 0 to 9;
  • x2 is an integer from 2 to 9;
  • x3 is an integer from 1 to 5;
  • x4 is an integer from 0 to 3;
  • y2 is an integer from 2 to 9;
  • y3 is an integer from 1 to 5;
  • y4 is an integer from 0 to 3;
  • z is an integer from 2 to 6. In some embodiments, z is 2, 4, or 5. In some embodiments, x0 and y0 are independently 2 to 6. In some embodiments, x0 and y0 are independently 4 or 5. In some embodiments, x1 and y1 are independently 2 to 6. In some embodiments, x1 and y1 are independently 4 or 5. In some embodiments, x2 and y2 are independently an integer from 2 to 8. In some embodiments, x2 and y2 are independently 3, 5, or 7. In some embodiments, x3 and y3 are both 1. In some embodiments, x4 and y4 are independently 0 or 1.
  • R 1 and R 2 are independently straight C 6 -C 10 alkyl, or -R 7 -CH (R 8 ) (R 9 ) , wherein R 7 is C 0 -C 5 alkylene, and R 8 and R 9 are independently C 2 -C 10 alkyl or C 2 -C 10 alkenyl.
  • the compound is a compound of formula (02-VI-A) , (02-VI-B) , (02-VI-C) , (02-VI-D) , (02-VI-E) , or (02-VI-F) :
  • z is an integer from 2 to 12;
  • y is an integer from 2 to 12;
  • x0 is an integer from 1 to 11;
  • x1 is an integer from 0 to 9;
  • x2 is an integer from 2 to 5;
  • x3 is an integer from 1 to 5;
  • x4 is an integer from 0 to 3;
  • z is an integer from 2 to 6. In some embodiments, z is 2, 4, or 5. In some embodiments, x0 is 4 or 5. In some embodiments, x1 is 4 or 5. In some embodiments, x2 is an integer from 2 to 5. In some embodiments, x2 is 3 or 5. In some embodiments, x3 is 0 or 1. In some embodiments, y is an integer from 2 to 6. In some embodiments, y is 5.
  • R 1 is straight C 6 -C 10 alkyl or -R 7 -CH (R 8 ) (R 9 ) , wherein R 7 is C 0 -C 5 alkylene, and R 8 and R 9 are independently C 2 -C 10 alkyl or C 2 -C 10 alkenyl.
  • R 2 and R f are each independently straight C 6 -C 18 alkyl, C 6 -C 18 alkenyl, or -R 7 -CH (R 8 ) (R 9 ) , wherein R 7 is C 0 -C 5 alkylene, and R 8 and R 9 are independently C 2 -C 10 alkyl or C 2 -C 10 alkenyl.
  • R d and R e are each independently H.
  • the compound is a compound in Table 2, or a pharmaceutically acceptable salt, prodrug, or stereoisomer thereof.
  • the cationic lipid described herein is a cationic lipid described in International Patent Publication No. WO 2022/152109, the entirety of which is incorporated herein by reference.
  • the cationic lipid is a compound of Formula (03-I) :
  • G 1 and G 2 are each independently a bond, C 2 -C 12 alkylene, or C 2 -C 12 alkenylene, wherein one or more -CH 2 -in G 1 and G 2 is optionally replaced by -O-;
  • R 1 and R 2 are each independently C 6 -C 24 alkyl or C 6 -C 24 alkenyl
  • R a , R b , R d , and R e are each independently H, C 1 -C 24 alkyl, or C 2 -C 24 alkenyl;
  • R c and R f are each independently C 1 -C 24 alkyl or C 2 -C 24 alkenyl
  • G 3 is C 2 -C 12 alkylene or C 2 -C 12 alkenylene, wherein part or all of alkylene or alkenylene is optionally replaced by C 3 -C 8 cycloalkylene, C 3 -C 8 cycloalkenylene, C 3 -C 8 cycloalkynylene, 4-to 8-membered heterocyclylene, C 6 -C 10 arylene, or 5-to 10-membered heteroarylene;
  • R 3 is hydrogen, C 1 -C 12 alkyl, C 2 -C 12 alkenyl, C 2 -C 12 alkynyl, C 3 -C 8 cycloalkyl, C 3 -C 8 cycloalkenyl, C 3 -C 8 cycloalkynyl, 4-to 8-membered heterocyclyl, C 6 -C 10 aryl, or 5-to 10-membered heteroaryl; or R 3 and G 1 , or part of G 1 , together with the nitrogen to which they are attached form a cyclic moiety; or R 3 and G 3 or part ofG 3 , together with the nitrogen to which they are attached form a cyclic moiety;
  • R 4 is C 1 -C 12 alkyl or C 3 -C 8 cycloalkyl
  • x 0, 1, or 2;
  • n I or 2;
  • n 1 or 2;
  • alkyl, alkenyl, alkynyl, cycloalkyl, cycloalkenyl, cycloalkynyl, heterocyclyl, aryl, heteroaryl, alkylene, alkenylene, cycloalkylene, cycloalkenylene, cycloalkynylene, heterocyclylene, arylene, heteroarylene, and cyclic moiety is independently optionally substituted.
  • the cationic lipid is a compound of Formula (03-II-A) :
  • the cationic lipid is a compound of Formula (03-II-B) :
  • the cationic lipid is a compound of Formula (03-II-C) :
  • the cationic lipid is a compound of Formula (03-II-D) :
  • G 1 and G 2 are each independently C 2 -C 12 alkylene. In some embodiments, G 1 and G 2 are each independently C 5 alkylene. In some embodiments, G 3 is C 2 -C 6 alkylene.
  • R 3 is C 1 -C 12 alkyl, C 2 -C 12 alkenyl, or C 3 -C 8 cycloalkyl. In some embodiments, R 3 is C 3 -C 8 cycloalkyl. In some embodiments, R 3 is unsubstituted. In some embodiments, R 4 is substituted C 1 -C 12 alkyl. In some embodiments, R 4 is-CH 2 CH 2 OH.
  • R 1 , R 2 , R c , and R f are each independently straight C 6 -C 18 alkyl, straight C 6 -C 18 alkenyl, or -R 7 -CH (R 8 ) (R 9 ) , wherein R 7 is C 0 -C 5 alkylene, and R 8 and R 9 are independently C 2 -C 10 alkyl or C 2 -C 10 alkenyl.
  • R 1 , R 2 , R c , and R f are each independently straight C 7 -C 15 alkyl, straight C 7 -C 15 alkenyl, or-R 7 -CH (R 8 ) (R 9 ) , wherein R 7 is C 0 -C 1 alkylene, and R 8 and R 9 are independently C 4 -C 8 alkyl or C 6 -C 10 alkenyl.
  • R a , R b , R d , and R e are each independently H.
  • the cationic lipid is a compound in Table 3, or a pharmaceutically acceptable salt, prodrug, or stereoisomer thereof.
  • the cationic lipid is a cationic lipid described in International Patent Application No. PCT/CN2022/094227, the entirety of which is incorporated herein by reference.
  • the cationic lipid is a compound of Formula (04-I) :
  • G 1 and G 2 are each independently a bond, C 2 -C 12 alkylene, or C 2 -C 12 alkenylene;
  • R 1 and R 2 are each independently C 5 -C 32 alkyl or C 5 -C 32 alkenyl
  • R a , R b , R d , and R e are each independently H, C 1 -C 24 alkyl, or C 2 -C 24 alkenyl;
  • R c and R f are each independently C 1 -C 32 alkyl or C 2 -C 32 alkenyl
  • R 0 is C 1 -C 12 alkyl, C 2 -C 12 alkenyl, C 3 -C 8 cycloalkyl, C 3 -C 8 cycloalkenyl, C 6 -C 10 aryl, or 4-to 8-membered heterocycloalkyl;
  • G 3 is C 2 -C 12 alkylene or C 2 -C 12 alkenylene
  • R 4 is C 1 -C 12 alkyl, C 2 -C 12 alkenyl, C 3 -C 8 cycloalkyl, C 3 -C 8 cycloalkenyl, C 6 -C 10 aryl, or 4-to 8-membered heterocycloalkyl;
  • R 5 is C 1 -C 12 alkyl, C 3 -C 8 cycloalkyl, C 3 -C 8 cycloalkenyl, C 6 -C 10 aryl, or 4-to 8-membered heterocycloalkyl;
  • x 0, 1, or 2;
  • s is 0 or 1;
  • each alkyl, alkenyl, cycloalkyl, cycloalkenyl, heterocycloalkyl, aryl, alkylene, alkenylene, arylene, and heteroarylene, is independently optionally substituted.
  • the cationic lipid is a compound of Formula (04-III) :
  • R 1 and R 2 are each independently C 5 -C 32 alkyl or C 5 -C 32 alkenyl
  • R 0 is C 1 -C 12 alkyl, C 2 -C 12 alkenyl, C 3 -C 8 cycloalkyl, C 3 -C 8 cycloalkenyl, C 6 -C 10 aryl, or 4-to 8-membered heterocycloalkyl;
  • G 3 is C 2 -C 12 alkylene or C 2 -C 12 alkenylene
  • G4 is C 2 -C 12 alkylene or C 2 -C 12 alkenylene
  • R 3 is -N (R 4 ) R 5 or -OR 6 ;
  • R 4 is C 1 -C 12 alkyl, C 2 -C 12 alkenyl, C 3 -C 8 cycloalkyl, C 3 -C 8 cycloalkenyl, C 6 -C 10 aryl, or 4-to 8-membered heterocycloalkyl;
  • R 5 is C 1 -C 12 alkyl, C 3 -C 8 cycloalkyl, C 3 -C 8 cycloalkenyl, C 6 -C 10 aryl, or 4-to 8-membered heterocycloalkyl; or R 4 and R 5 , together with the nitrogen to which they are attached form a cyclic moiety;
  • R 6 is hydrogen, C 1 -C 12 alkyl, C 3 -C 8 cycloalkyl, C 3 -C 8 cycloalkenyl, or C 6 -C 10 aryl; and wherein each alkyl, alkenyl, cycloalkyl, cycloalkenyl, heterocycloalkyl, aryl, alkylene, alkenylene, and cyclic moiety is independently optionally substituted.
  • the cationic lipid is a compound of Formula (04-IV) :
  • G 3 is C 2 -C 4 alkylene. In some embodiments, G4 is C 2 -C 4 alkylene.
  • R 0 is C 1 -C 6 alkyl.
  • R 3 is -OH.
  • R 3 is -N (R 4 ) R 5 .
  • R 4 is C 3 -C 8 cycloalkyl.
  • R 4 is unsubstituted.
  • R 5 is -CH 2 CH 2 OH.
  • R 1 and R 2 are each independently branched C 6 -C 24 alkyl or branched C 6 -C 24 alkenyl.
  • R 1 and R 2 are each independently -R 7 -CH (R 8 ) (R 9 ) , wherein R 7 is C 1 -C 5 alkylene, and R 8 and R 9 are independently C 2 -C 10 alkyl or C 2 -C 10 alkenyl.
  • R 1 is straight C 6 -C 24 alkyl and R 2 is branched C 6 -C 24 alkyl.
  • R 1 is straight C 6 -C 24 alkyl and R 2 is -R 7 -CH (R 8 ) (R 9 ) , wherein R 7 is C 1 -C 5 alkylene, and R 8 and R 9 are independently C 2 -C 10 alkyl.
  • the cationic lipid is a compound in Table 4, or a pharmaceutically acceptable salt, prodrug, or stereoisomer thereof.
  • the cationic lipid contained in the particles or compositions provided herein is a cationic lipid described in U.S. Patent Nos. US 10442756B2, US9868691B2, or US9868692B2, all of which are incorporated herein by reference.
  • the cationic lipid is a compound Formula (05-I) :
  • 1 is selected from 1, 2, 3, 4, and 5;
  • n is selected from 5, 6, 7, 8, and 9;
  • M 1 is a bond or M′
  • M and M′ are independently selected from -C (O) O-, -OC (O) -, -C (O) N (R′) -, -P (O) (OR′) O-, -S-S-, an aryl group, and a heteroaryl group; and
  • R 2 and R 3 are both C 1 -C14 alkyl, or C 2 -C14 alkenyl
  • R 8 is selected from the group consisting of C 3 -C 6 carbocycle and heterocycle
  • R 9 is selected from the group consisting of H, CN, NO 2 , C 1 -C 6 alkyl, -OR, -S (O) 2 R, -S (O) 2 N (R) 2 , C 2 -C 6 alkenyl, C 3 -C 6 carbocycle and heterocycle;
  • each R is independently selected from the group consisting of C 1 -C 3 alkyl, C 2 -C 3 alkenyl, and H;
  • R′ is a linear alkyl
  • the cationic lipid is SM102 or Lipid 5:
  • the cationic lipid is a cationic lipid described in U.S. Patent No. US 10166298B2, the entire teachings of which are incorporated herein by reference.
  • the cationic lipid is a compound of Formula (06-I) :
  • G 1 and G 2 are each independently unsubstituted C 1 -C 12 alkylene or C 1 -C 12 alkenylene;
  • G 3 is C 1 -C 24 alkylene, C 1 -C 24 alkenylene, C 3 -C 8 cycloalkylene, C 3 -C 8 cycloalkenylene;
  • R a is H or C 1 -C 12 alkyl
  • R 1 and R 2 are each independently C 6 -C 24 alkyl or C 6 -C 24 alkenyl
  • R 4 is C 1 -C 12 alkyl
  • R 5 is H or C 1 -C 6 alkyl
  • the cationic lipid is a compound of Table 5, or a pharmaceutically acceptable salt, prodrug, or stereoisomer thereof.
  • the cationic lipids of the present disclosure are the same as those disclosed in International Application Publication No. WO 2010/144740, the entire teachings of which are incorporated herein by reference.
  • the cationic lipid is a compound represented by Formula (07-I) , also named as compound 07-I:
  • the ionizable lipid used in the LNPs according to the present invention is selected from
  • y and z are each independently an integer from 4 to 6,
  • s is an integer from 2 to 4,
  • t is an integer from 1 to 3
  • R 1 and R 2 are each independently C 12 -C 22 alkyl
  • R 4 is C 3 -C 8 cycloalkyl
  • R 6 is hydrogen or hydroxyl
  • 1 is selected from 1, 2, 3, 4, and 5;
  • n is selected from 5, 6, 7, 8, and 9;
  • M 1 is -C (O) O-;
  • R 4 is - (CH 2 ) n OH, and n is selected from 1, 2, 3, 4, or 5;
  • M is -OC (O) -
  • R 2 and R 3 are both C 6-10 alkyl
  • G 1 and G 2 are each independently unsubstituted C 4 -C 8 alkylene
  • G 3 is C 3 -C 8 alkylene
  • R 1 and R 2 are each independently C 12 -C 22 alkyl
  • R 3 is H or OH
  • R 1 and R 2 are each independently C 6 -C 24 alky
  • R 3 is-OR 6 ;
  • R 6 is hydrogen
  • z is an integer from 2 to 12;
  • x1 is an integer from 0 to 9;
  • y1 is an integer from 0 to 9;
  • R 1 and R 2 are each independently C 6 -C 24 alkyl or C 6 -C 24 alkenyl
  • R 3 is -OR 6 ;
  • R 6 is hydrogen
  • z is an integer from 2 to 12;
  • y is an integer from 2 to 12;
  • x1 is an integer from 2 to 5;
  • R 1 and R 2 are each independently C 6 -C 24 alkyl
  • R 3 is -OR 6 ;
  • R 6 is hydrogen
  • z is an integer from 2 to 12;
  • x2 is an integer from 2 to 9;
  • x4 is an integer from 0 to 3;
  • y2 is an integer from 2 to 9;
  • y4 is an integer from 0 to 3
  • G 1 and G 2 are each independently C 3 -C 8 alkylene
  • R 1 is independently C 6 -C 24 alkyl
  • R 2 is independently C 6 -C 24 alkyl
  • G 3 is C 2 -C 12 alkylene
  • R 3 is C 3 -C 8 cycloalkyl
  • R 4 is C 1 -C 4 hydroxylalkyl
  • n 1 or 2;
  • n 1 or 2
  • the ionizable lipid used in the LNPs according to the present invention is selected from the following compounds:
  • the LNP comprises a polymer conjugated lipid.
  • Polymers that can be incorporated into polymer conjugated lipids include polyamines, polyethers, polyamides, polyesters, polycarbamates, polyureas, polycarbonates, polystyrenes, polyimides, polysulfones, polyurethanes, polyacetylenes, polyethylenes, polyethyleneimines, polyisocyanates, polyacrylates, polymethacrylates, polyacrylonitriles, and polyarylates.
  • a polymer may include poly (caprolactone) (PCL) , ethylene vinyl acetate polymer (EVA) , poly (lactic acid) (PLA) , poly (L-lactic acid) (PLLA) , poly (glycolic acid) (PGA) , poly (lactic acid-co-glycolic acid) (PLGA) , poly (L-lactic acid-co-glycolic acid) (PLLGA) , poly (D, L-lactide) (PDLA) , poly (L-lactide) (PLLA) , poly (D, L-lactide-co-caprolactone) , poly (D, L-lactide-co-caprolactone-co-glycolide) , poly (D, L-lactide-co-PEO-co-D, L-lactide) , poly (D, L-lactide-co-PPO-co-D, L-lactide) , polyalkyl cyanoacralate, poly (
  • the polymer conjugated lipid is a PEGylated lipid (PEG lipids) .
  • PEG lipids PEGylated lipids
  • a polymer conjugated lipid component in an LNP can improve colloidal stability and/or reduce protein absorption of the nanoparticles.
  • Exemplary PEGylated lipids that can be used in connection with the present disclosure include but are not limited to PEG-modified phosphatidylethanolamines, PEG-modified phosphatidic acids, PEG-modified ceramides, PEG-modified dialkylamines, PEG-modified diacylglycerols, PEG-modified dialkylglycerols, and mixtures thereof.
  • the PEG-conjugated lipid may be a PEG-modifiedphosphatidylethanolamine, PEG-modified phosphatidic acid, PEG-modified ceramide, PEG-modified dialkylamine, PEG-modified diacylglycerol, or a PEG-modified dialkylglycerol.
  • the PEG lipid may be PEG-c-DOMG, PEG-DMG, PEG-DLPE, PEG-DMPE, PEG-DPPC, PEG-DSPE, Ceramide-PEG2000, or ChoI-PEG2000.
  • the PEGylated lipid is a PEGylated diacylglycerol (PEG-DAG) such as 1- (monomethoxy-polyethyleneglycol) -2, 3-dimyristoylglycerol (PEG-DMG) , a pegylated phosphatidylethanoloamine (PEG-PE) , a PEG succinate diacylglycerol (PEG-S-DAG) such as 4-O- (2’, 3’-di (tetradecanoyloxy) propyl-1-O- ( ⁇ -methoxy (polyethoxy) ethyl) butanedioate (PEG-S-DMG) , a pegylated ceramide (PEG-cer) , or a PEG dialkoxypropylcarbamate such as ⁇ -methoxy (polyethoxy) ethyl-N- (2, 3-di (tetradecanoxy) prop
  • the PEGylated lipid is present in a concentration ranging from 1.0 to 2.5 molar percent. In some embodiments, the polymer conjugated lipid is present in a concentration of about 1.7 molar percent. In some embodiments, the polymer conjugated lipid is present in a concentration of about 1.5 molar percent.
  • the molar ratio of the ionizable lipid to the polymer conjugated lipid ranges from about 20 ⁇ 1 to about 100 ⁇ 1. In some embodiments, the molar ratio of the ionizable lipid to polymer conjugated lipid ranges from about 25 ⁇ 1 to about 80 ⁇ 1 . In some embodiments, the molar ratio of the ionizable lipid to polymer conjugated lipid ranges from about 30 ⁇ 1 to about 60 ⁇ 1. In some embodiments, the molar ratio of the ionizable lipid to polymer conjugated lipid ranges from about 30 ⁇ 1 to about 50 ⁇ 1.
  • the PEGylated lipid has the following Formula:
  • R 12 and R 13 are each independently a straight or branched, alkyl or alkenyl chain containing from 10 to 30 carbon atoms, wherein the alkyl chain is optionally interrupted by one or more ester bonds;
  • w has a mean value ranging from 30 to 60.
  • R 12 and R 13 are each independently straight, saturated alkyl chains containing from 12 to 16 carbon atoms.
  • the average w ranges from 42 to 55, for example, the average w is 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54 or 55. In some embodiments, the average w is about 49.
  • the PEGylated lipid has the following Formula:
  • the LNPs comprise a lipid stabilizer.
  • the lipid stabilizer comprises a sterol.
  • the lipid stabilizer comprises a corticosteroid.
  • the lipid stabilizer comprises two or more components.
  • the lipid stabilizer comprises a corticosteroid and a sterol.
  • the sterol is selected from the group consisting of cholesterol, fecosterol, sitosterol, ergosterol, campesterol, stigmasterol, and brassicasterol.
  • the corticosteroid is selected from the group consisting ofprednisolone, dexamethasone, prednisone, and hydrocortisone.
  • the lipid stabilizer comprises tomatidine, tomatine, ursolic acid, or alpha-tocopherol.
  • the lipid stabilizer comprises one or more compounds selected from the group consisting of cholesterol, fecosterol, sitosterol, ergosterol, campesterol, stigmasterol, brassicasterol, prednisolone, dexamethasone, prednisone, hydrocortisone, tomatidine, tomatine, ursolic acid, and alpha-tocopherol.
  • the lipid stabilizer is cholesterol.
  • the LNPs disclosed herein further comprise a therapeutic payload.
  • the payload can be any substance or compound that has a therapeutic or prophylactic effect.
  • the therapeutic payload is a small molecule, a cytotoxin, a radioactive ion, a chemotherapeutic compound, a vaccine, or a compound that elicits an immune response.
  • the LNPs disclosed herein comprise a nucleic acid.
  • the nucleic acid is a DNA.
  • the DNA is catalytic DNA, plasmid DNA, aptamer, or complementary DNA (cDNA) .
  • the nucleic acid is an RNA.
  • the RNA is a messenger RNA (mRNA) , antisense oligonucleotide, microRNA (miRNA) , miRNA inhibitor (e.g., antagomir or antimir) , messenger-RNA-interfering complementary RNA (micRNA) , multivalent RNA, dicer substrate RNA (dsRNA) , small hairpin RNA (shRNA) , antisense RNA, transfer RNA (tRNA) , asymmetrical interfering RNA (aiRNA) , a ribozyme, an aptamer, or a vector.
  • the RNA is an mRNA hybrid.
  • the nucleic acid is an mRNA.
  • the mRNA encodes a protein.
  • the protein is an antibody.
  • the antibody is a bispecific antibody.
  • the LNPs comprise an RNAi agent or RNAi-inducing agent.
  • the weight ratio of the ionizable lipid to the therapeutic payload is from 5 ⁇ 1 to 20 ⁇ 1.
  • compositions comprising Lipid Nanoparticles (LNPs)
  • the present disclosure is inclusive with respect to compositions comprising the LNPs described herein.
  • the composition comprises a plurality of LNPs.
  • the plurality of LNPs have a polydispersity index (PDI) of 0.001 to 0.2.
  • the plurality of LNPs have a polydispersity index (PDI) of 0.001 to 0.1.
  • the LNPs have a PDI of 0.005 to 0.05.
  • some of the LNPs of the LNP composition comprise mRNA.
  • the mRNA encapsulation efficiency (EE%-i.e., the percentage of individual LNPs in the composition that encapsulate the mRNA) of the LNP composition is from 70%to 100%.
  • the EE%of the LNP composition is from 80 to 95%.
  • the EE%of the LNP composition is from 85 to 95%.
  • the EE%of the LNP composition is from 90 to 95%.
  • the EE% is 80%or greater.
  • the EE% is 85%or greater.
  • the LNP composition is able to increase protein expression compared to comparable LNP compositions that do not comprise a steroid-containing phospholipid.
  • any of the molar percentages or molar ratios described above for the sterol-containing phospholipid, the ionizable lipid, the polymer conjugated lipid, and the lipid stabilizer may be combined with each other in any embodiment describing the composition of LNPs described herein, such that every combination is contemplated as though each and every combination were specifically and individually disclosed.
  • the LNPs here can be made according to the methods that are well known in the art.
  • the method comprises solubilizing the lipid components (e.g., a phospholipid, an ionizable lipid, a polymer conjugated lipid, and a lipid stabilizer) in a solvent.
  • the method comprises the steps:
  • lipid components e.g., a phospholipid, an ionizable lipid, a polymer conjugated lipid, and optionally a lipid stabilizer
  • the solvent in step (a) is a polar solvent. In some embodiments, the solvent in step (a) is an alcohol solvent. In some embodiments, the solvent in step (a) is methanol, ethanol, n-propanol, or isopropanol. In some embodiments, the solvent in step (a) is ethanol.
  • the solvent in step (b) is an aqueous solvent. In some embodiments, the solvent in step (b) is an aqueous buffer. In some embodiments, the solvent in step (b) is a citrate buffer. In some embodiments, the citrate buffer has a citrate concentration of 5 to 100 mM.In some embodiments, the citrate buffer has a citrate concentration of 10 to 50 mM. In some embodiments, the aqueous solvent of step (b) has a pH of 2-6. In some embodiments, the aqueous solvent of step (b) has a pH of 3-5.
  • the mixing of step (c) occurs with a weight ratio of lipid ⁇ mRNA of 10 ⁇ 1 to 30 ⁇ 1. In some embodiments, the mixing of step (c) occurs at a volume ratio of lipid ⁇ mRNA of 1 ⁇ 1 to 1 ⁇ 5. In some embodiments, the volume ratio is 1 ⁇ 2 to 1 ⁇ 4. In some embodiments, the volume ratio is about 1 ⁇ 3. In some embodiments, the mixing of step (c) is performed with a microfluidic apparatus. In some embodiments, the microfluidic apparatus has a flow rate of 9 to 30 mL/min.
  • the mRNA mixture has an mRNA concentration from 1 to 3, 5, 7, 10, 12, 15, 20, 30, 40 or 50 mM. In some embodiments, the concentration is from 3 to 5, 7, 10, 12, 15, 20, or 30mM. In some embodiments, the concentration is from 5 to 7, 10, 12, 15, 20, or 30 mM. In some embodiments, the concentration is from 7 to 10, 12, 15, 20, or 30 mM. In some embodiments, the concentration is from 10 to 12, 15, 20, or 30 mM. In some embodiments, the concentration is from 12 to 15, 20, or 30 mM. In some embodiments, the concentration is from 15 to 20 or 30 mM. In some embodiments, the concentration is from 20 to 30 mM. In some embodiments, the concentration is about 5, 7, 10, 12, 15, or 20 mM.
  • the concentration is about 5, 7, 10, 12, or 15 mM. In some embodiments, the concentration is about 10 mM. In some embodiments, the concentration is about 12 mM. In some embodiments, the concentration is about 15 mM. In some embodiments, the concentration is about 20 mM.
  • the method further comprises a step (d) :
  • the sterile filter is a 0.2 ⁇ m sterile filter.
  • the present disclosure is inclusive with respect to potentially useful methods that make use of the LNPs described herein.
  • a method for expressing protein in a cell comprising introducing an LNP or composition thereof, as described above, to a cell.
  • the cell is a mammalian cell.
  • the LNP or composition thereof is administered systemically to a mammal.
  • the mammal is a human.
  • the LNP composition is able to increase protein expression compared to comparable LNP compositions that do not comprise a steroid-containing phospholipid.
  • Kits comprising a phospholipid containing a sterol moiety
  • kits comprising a phospholipid containing a sterol moiety and packaging for said phospholipid.
  • the kit further comprises an ionizable lipid.
  • the kit further comprises a polymer conjugated lipid.
  • the kit further comprises a lipid stabilizer.
  • the kit further comprises an ionizable lipid, a polymer conjugated lipid, and a lipid stabilizer.
  • the ionizable lipid is a cationic lipid.
  • the polymer conjugated lipid is a PEGylated lipid.
  • the lipid stabilizer is cholesterol.
  • the kit further comprises a cationic lipid, a PEGylated lipid, and cholesterol.
  • any embodiment of the compounds provided herein, as set forth above, and any specific substituent and/or variable in the compound provided herein, as set forth above, may be independently combined with other embodiments and/or substituents and/or variables of the compounds to form embodiments not specifically set forth above.
  • substituents and/or variables may be listed for any particular group or variable, it is understood that each individual substituent and/or variable may be deleted from the particular embodiment and/or claim and that the remaining list of substituents and/or variables will be considered to be within the scope of embodiments provided herein.
  • Embodiment 1 A lipid nanoparticle (LNP) comprising
  • Embodiment 2 The LNP of embodiment 1, wherein the phospholipid has a structure selected from:
  • Embodiment 3 The LNP of embodiment 1 or 2, wherein the phospholipid has the structure:
  • Embodiment 4 The LNP of embodiment 1 or 2, wherein the phospholipid has the structure:
  • Embodiment 5 The LNP of any one of embodiments 1 to 4, wherein the LNP has a molar ratio of the ionizable lipid to the phospholipid from 20 ⁇ 1 to 2 ⁇ 1.
  • Embodiment 6 The LNP of embodiment 5, wherein the molar ratio of the ionizable lipid to the phospholipid is from 15 ⁇ 1 to 5 ⁇ 1.
  • Embodiment 7 The LNP of any one of embodiments 1 to 4, wherein the ionizable lipid comprises from 40 to 80 mol%of a total amount of lipids in the LNP.
  • Embodiment 8 The LNP of embodiment 7, wherein the ionizable lipid comprises from 50 to 70 mol%of the total amount of lipids in the LNP.
  • Embodiment 9 The LNP of any one of embodiments 1 to 8, wherein the ionizable lipid is a cationic lipid.
  • Embodiment 10 The LNP of any one of embodiments 1 to 8, wherein the ionizable lipid is a compound according to any one of the formula selected from 01-I, 01-II, 02-I, 02-II, 03-I, 03-II-A, 03-II-B, 03-II-C, 03-II-D, 04-I, 04-III, 04-IV, 05-I, 06-I, and sub-formulas thereof, or wherein the ionizable lipid is a cationic lipid selected from the compounds listed in any one of Tables 1 to 5.
  • Embodiment 11 The LNP of any one of embodiments 1 to 10, wherein the polymer conjugated lipid comprises from 1 to 2%of a total amount of lipids in the LNP.
  • Embodiment 12 The LNP of embodiment 11, wherein the polymer conjugated lipid comprises 1.5%of the total amount of lipids in the LNP.
  • Embodiment 13 The LNP of any one of embodiments 1 to 12, wherein the LNP has a molar ratio of the polymer conjugated lipid to the phospholipid from 1 ⁇ 5 to 1 ⁇ 10.
  • Embodiment 14 The LNP of any one of embodiments 1 to 13, wherein the polymer conjugated lipid is a PEGylated lipid.
  • Embodiment 15 The LNP of any one of embodiments 1 to 13, wherein the polymer conjugated lipid is a PEGylated lipid with the structure:
  • R 12 and R 13 are each independently a straight or branched, alkyl or alkenyl chain containing from 10 to 30 carbon atoms, wherein the alkyl chain is optionally interrupted by one or more ester bonds;
  • w is an integer ranging from 30 to 60.
  • Embodiment 16 The LNP of any one of embodiments 1 to 15, wherein the polymer conjugated lipid is a PEGylated lipid with the structure:
  • w is an integer ranging from 30 to 60.
  • Embodiment 17 The LNP of embodiment 15 or 16, wherein w is an integer ranging from 45 to 55.
  • Embodiment 18 The LNP of embodiment 15 or 16, wherein w is about 49.
  • Embodiment 19 The LNP of any one of embodiments 1 to 14, wherein the polymer conjugated lipid is DMG-PEG or DMPE-PEG.
  • Embodiment 20 The LNP of any one of embodiments 1 to 19, further comprising a lipid stabilizer.
  • Embodiment 21 The LNP of embodiment 20, wherein the LNP has a molar ratio of the lipid stabilizer to the phospholipid from 10 ⁇ 1 to 1 ⁇ 4.
  • Embodiment 22 The LNP of embodiment 21, wherein the molar ratio of the lipid stabilizer to the phospholipid is from 5 ⁇ 1 to 1 ⁇ 1.
  • Embodiment 23 The LNP of embodiment 21, wherein the molar ratio of the lipid stabilizer to the phospholipid is from 4 ⁇ 1 to 3 ⁇ 1.
  • Embodiment 24 The LNP of any one of embodiments 20 to 23, wherein the lipid stabilizer comprises from 5 to 50 mol%of a total amount of lipids in the LNP.
  • Embodiment 25 The LNP of embodiment 24, wherein the lipid stabilizer comprises from 8 to 40 mol%of the total amount of lipids in the LNP.
  • Embodiment 26 The LNP of embodiment 24, wherein the lipid stabilizer comprises from 10 to 30 mol%of the total amount of lipids in the LNP.
  • Embodiment 27 The LNP of any one of embodiments 1 to 26, wherein the phospholipid comprises from 1 to 30 mol%of a total amount of lipids in the LNP.
  • Embodiment 28 The LNP of embodiment 27, wherein the phospholipid comprises from 2 to 25 mol%of the total amount of lipids in the LNP.
  • Embodiment 29 The LNP of embodiment 27, wherein the phospholipid comprises from 3 to 20 mol%of the total amount of lipids in the LNP.
  • Embodiment 30 The LNP of embodiment 27, wherein the phospholipid comprises from 5 to 15 mol%of the total amount of lipids in the LNP.
  • Embodiment 31 The LNP of embodiment 27, wherein the phospholipid comprises about 10 mol%of the total amount of lipids in the LNP.
  • Embodiment 32 The LNP of any one of embodiments 1 to 31, wherein the LNP has a size from 50 nm to 150 nm, as determined using dynamic light scattering.
  • Embodiment 33 The LNP of embodiment 32, wherein the size is from 60 nm to 140 nm.
  • Embodiment 34 The LNP of embodiment 32, wherein the size is from 80 nm to 100 nm.
  • Embodiment 35 The LNP of embodiment 32, wherein the size is from 85 nm to 95 nm.
  • Embodiment 36 The LNP of any one of embodiments 1 to 35, wherein the LNP encapsulates mRNA.
  • Embodiment 37 A composition comprising lipid nanoparticles (LNPs) , wherein each LNP is an LNP of any one of embodiments 1 to 36.
  • LNPs lipid nanoparticles
  • Embodiment 38 The composition of embodiment 37, wherein at least 80%of the LNPs encapsulate mRNA.
  • Embodiment 39 The composition of embodiment 37, wherein at least 85%of the LNPs encapsulate mRNA.
  • Embodiment 40 A method for expressing protein in a cell, comprising introducing the LNP of embodiment 36 or the composition of embodiment 38 or 39, to the cell.
  • Embodiment 41 The method of embodiment 40, wherein the cell is a mammalian cell.
  • Embodiment 42 A method for delivering a protein to a subject, comprising administering the LNP of embodiment 36 or the composition of embodiment 38 or 39 to the individual, wherein the mRNA encodes the protein.
  • Embodiment 43 The method of embodiment 42, wherein the LNP or the composition is administered systemically.
  • Embodiment 44 The method of embodiment 42, wherein the subject is a mammal.
  • Embodiment 45 The method of embodiment 42, wherein the subject is a human.
  • Embodiment 46 A lipid nanoparticle (LNP) comprising a phospholipid, wherein the phospholipid has a structure selected from:
  • Embodiment 47 The LNP of embodiment 46, wherein the phospholipid has the structure:
  • Embodiment 48 The LNP of embodiment 46, wherein the phospholipid has the structure:
  • Embodiment 49 The LNP of any one of embodiments 46-48, further comprising an ionizable lipid.
  • Embodiment 50 The LNP of embodiment 49, wherein the LNP has a molar ratio of the ionizable lipid to the phospholipid from 20 ⁇ 1 to 2 ⁇ 1.
  • Embodiment 51 The LNP of embodiment 50, wherein the molar ratio of ionizable lipid to phospholipid is from 15 ⁇ 1 to 5 ⁇ 1.
  • Embodiment 52 The LNP of embodiment 49, wherein the ionizable lipid comprises from 40 to 80 mol%of a total amount oflipids in the LNP.
  • Embodiment 53 The LNP of embodiment 52, wherein the ionizable lipid comprises from 50 to 70 mol%of the total amount of lipids in the LNP.
  • Embodiment 54 The LNP of any one of embodiments 49 to 53, wherein the ionizable lipid is a compound according to any one of the formula selected from 01-I, 01-II, 02-I, 02-II, 03-I, 03-II-A, 03-II-B, 03-II-C, 03-II-D, 04-I, 04-III, 04-IV, 05-I, 06-I, and sub-formulas thereof, or wherein the ionizable lipid is a cationic lipid selected from the compounds listed in any one of Tables 1 to 5.
  • Embodiment 55 The LNP of any one of embodiments 49 to 54, wherein the ionizable lipid is a cationic lipid.
  • Embodiment 56 The LNP of any one of embodiments 46 to 55, further comprising a polymer conjugated lipid.
  • Embodiment 57 The LNP of embodiment 56, wherein the polymer conjugated lipid comprises from 1 to 2%of a total amount oflipids in the LNP.
  • Embodiment 58 The LNP of embodiment 57, wherein the polymer conjugated lipid comprises 1.5%of the total amount of lipids in the LNP.
  • Embodiment 59 The LNP of any one of embodiments 46 to 58, wherein the LNP has a molar ratio of the polymer conjugated lipid to the phospholipid from 1 ⁇ 5 to 1 ⁇ 10.
  • Embodiment 60 The LNP of any one of embodiments 56 to 59, wherein the polymer conjugated lipid is a PEGylated lipid.
  • Embodiment 61 The LNP of any one of embodiments 56 to 60, wherein the polymer conjugated lipid is a PEGylated lipid with the structure:
  • R 12 and R 13 are each independently a straight or branched, alkyl or alkenyl chain containing from 10 to 30 carbon atoms, wherein the alkyl chain is optionally interrupted by one or more ester bonds;
  • w is an integer ranging from 30 to 60.
  • Embodiment 62 The LNP of any one of embodiments 56 to 61, wherein the polymer conjugated lipid is a PEGylated lipid with the structure:
  • w is an integer ranging from 30 to 60.
  • Embodiment 63 The LNP of embodiment 61 or 62, wherein w is an integer ranging from 45 to 55.
  • Embodiment 64 The LNP of embodiment 61 or 62, wherein w is about 49.
  • Embodiment 65 The LNP of any one of embodiments 56 to 60, wherein the polymer conjugated lipid is DMG-PEG or DMPE-PEG.
  • Embodiment 66 The LNP of any one of embodiments 46 to 65, further comprising a lipid stabilizer.
  • Embodiment 67 The LNP of embodiment 66, wherein the LNP has a molar ratio of the lipid stabilizer to the phospholipid from 10 ⁇ 1 to 1 ⁇ 4.
  • Embodiment 68 The LNP of embodiment 67, wherein the molar ratio of the lipid stabilizer to the phospholipid is from 5 ⁇ 1 to 1 ⁇ 1.
  • Embodiment 69 The LNP of embodiment 67, wherein the molar ratio of the lipid stabilizer to the phospholipid is from 4 ⁇ 1 to 3 ⁇ 1.
  • Embodiment 70 The LNP of any one of embodiments 66 to 69, wherein the lipid stabilizer comprises from 5 to 50 mol%of a total amount of lipids in the LNP.
  • Embodiment 71 The LNP of embodiment 70, wherein the lipid stabilizer comprises from 8 to 40 mol%of the total amount of lipids in the LNP.
  • Embodiment 72 The LNP of embodiment 70, wherein the lipid stabilizer comprises from 10 to 30 mol%of the total amount of lipids in the LNP.
  • Embodiment 73 The LNP of any one of embodiments 46 to 72, wherein the phospholipid comprises from 1 to 30 mol%of a total amount of lipids in the LNP.
  • Embodiment 74 The LNP of embodiment 73, wherein the phospholipid comprises from 2 to 25 mol%of the total amount of lipids in the LNP.
  • Embodiment 75 The LNP of embodiment 73, wherein the phospholipid comprises from 3 to 20 mol%of the total amount of lipids in the LNP.
  • Embodiment 76 The LNP of embodiment 73, wherein the phospholipid comprises from 5 to 15 mol%of the total amount of lipids in the LNP.
  • Embodiment 77 The LNP of embodiment 73, wherein the phospholipid comprises about 10 mol%of the total amount of lipids in the LNP.
  • Embodiment 78 The LNP of any one of embodiments 46 to 77, wherein the LNP has a size from 50 nm to 150 nm, as determined using dynamic light scattering.
  • Embodiment 79 The LNP of embodiment 78, wherein the size is from 60 nm to 140 nm.
  • Embodiment 80 The LNP of embodiment 78, wherein the size is from 80 nm to 100 nm.
  • Embodiment 81 The LNP of embodiment 78, wherein the size is from 85 nm to 95 nm.
  • Embodiment 82 The LNP of any one of embodiments 46 to 81, wherein the LNP encapsulates mRNA.
  • Embodiment 83 A composition comprising lipid nanoparticles (LNPs) , wherein each LNP is an LNP of any one of embodiments 46 to 82.
  • LNPs lipid nanoparticles
  • Embodiment 84 The composition of embodiment 83, wherein at least 80%of the LNPs encapsulate mRNA.
  • Embodiment 85 The composition of embodiment 83, wherein at least 85%of the LNPs encapsulate mRNA.
  • Embodiment 86 A method for expressing protein in a cell, comprising introducing the LNP of embodiment 82 or the composition of embodiment 84 or 85, to the cell.
  • Embodiment 87 The method of embodiment 86, wherein the cell is a mammalian cell.
  • Embodiment 88 A method for delivering a protein to a subject, comprising administering the LNP of embodiment 82 or the composition of embodiment 84 or 85 to the individual, wherein the mRNA encodes the protein.
  • Embodiment 89 The method of embodiment 88, wherein the LNP or the composition is administered systemically.
  • Embodiment 90 The method of embodiment 88, wherein the subject is a mammal.
  • Embodiment 91 The method of embodiment 88, wherein the subject is a human.
  • HPLC purification is carried out on a Waters 2767 equipped with a diode array detector (DAD) on an Inertsil Pre-C8 OBD column, generally with water containing 0.1%trifluoroacetic acid (TFA) as solvent A and acetonitrile as solvent B.
  • DAD diode array detector
  • TFA trifluoroacetic acid
  • LCMS analysis is conducted on a Shimadzu (LC-MS2020) System. Chromatography is performed on a SunFire C18, generally with water containing 0.1%formic acid as solvent A and acetonitrile containing 0.1%formic acid as solvent B.
  • OChemsPC refers to the following compound:
  • PChemsPC refers to the following compound:
  • DChemsPC refers to the following compound:
  • DSPC distearoylphosphatidylcholine.
  • compound 01-1 refers to compound 01-1 in Table 1.
  • Cho is an abbreviation for cholesterol.
  • DMG-PEG refers to 1, 2-dimyristoyl-rac-glycero-3-methoxypolyethylene glycol-2000.
  • ALC-0315 refers to compound 06-1, MC3 refers to compound 07-I.
  • Compound 02-1 was prepared according to the scheme below.
  • Compound 02-3 was prepared in analogous fashion as Compound 02-1, using corresponding starting material.
  • Compound 02-2 was prepared according to the scheme below.
  • Compound 02-4 was prepared according to the scheme below.
  • Compound 02-9 was prepared according to the scheme below.
  • Compound 02-14 was prepared in analogous fashion as Compound 02-9, using corresponding starting material.
  • Compound 02-10 was prepared according to the scheme below.
  • Compound 02-11 was prepared in analogous fashion as Compound 02-10, using corresponding starting material.
  • Compound 02-12 was prepared according to the scheme below.
  • Compound 02-20 was prepared according to the scheme below.
  • Compound 04-1 was prepared according to the scheme below.
  • Compound 04-2 was prepared according to the scheme below.
  • Compound 04-7 was prepared according to the scheme below.
  • Compound 04-8 was prepared according to the scheme below.
  • Compound 04-65 was prepared according to the scheme below.
  • Compound 04-68 was prepared according to the scheme below.
  • Example 15 Initial screening of LNPs comprising a steroid containing phospholipid
  • the specified amounts of the lipid components were solubilized in ethanol at the specified molar ratios (see Table 6) .
  • the LNPs were prepared at a total lipid to mRNA weight ratio of approximately 10: 1 to 30: 1 by mixing the ethanolic lipid solution with the aqueous mRNA solution at a volume ratio of 1: 3 using a microfluidic apparatus, total flow rate ranging from 9-30 mL/min. Ethanol was thereby removed and replaced by Dulbecco′sphosphate-buffered saline (DPBS) using dialysis. Finally, the lipid nanoparticles were filtered through a 0.2 ⁇ m sterile filter.
  • DPBS Dulbecco′sphosphate-buffered saline
  • Lipid nanoparticle size were determined by dynamic light scattering using a Malvern Zetasizer Nano ZS (Malvern UK) using a 173° backscatter detection mode.
  • the encapsulation efficiency of lipid nanoparticles was determined using a Quant-it Ribogreen RNA quantification assay kit (Thermo Fisher Scientific, UK) according to the manufacturer's instructions.
  • lipid nanoparticle formulations were diluted 20-fold in PBS and transferred 1 mL in measurement cuvette.
  • the LNP encapsulation efficiency (EE%) was determined using a Quant-it RiboGreen RNA assay kit, LNP formulations were diluted to 0.5 ⁇ g/mL in Tris-EDTA and 0.1%Triton respectively.
  • ribogreen reagent were diluted 200-fold with Tris-EDTA buffer and mix at the same volume as diluted LNP formulation.
  • Fluorescence intensity was measured at room temperature in a Molecular Devices Spectramax iD3 spectrometer using excitation and emission wavelengths of 488 nm and 525 nm. EE% was calculated based on the ratio of encapsulated to total RNA fluorescence intensity.
  • Lipid nanoparticles encapsulating human erythropoietin (hEPO) mRNA were prepared as described above, and systemically administered to 6-8 week old female ICR mice (Xipuer-Bikai, Shanghai) at 0.5mg/kg dose by tail vein injection. Mice were euthanized by CO 2 overdoses at 6 hours post administration, and blood samples were taken for hEPO measurement. Particularly, serum was separated from total blood by centrifugation at 5000g for 10 minutes at 4 °C, snap-frozen and stored at -80 °C for analysis. The serum hEPO level was measured using an ELISA assay carried out using a commercial kit (DEP00, R&D systems) according to manufacturer's instructions. The hEPO expression levels ( ⁇ g/ml) measured from the tested group are plotted in FIG. 1 and summarized in Table 6.
  • Example 16 Expression levels of loaded LNPs comprising a steroid containing phospholipid
  • LNPs containing 45-60 mol%compound 01-1 resulted in the highest protein expression level.
  • PChemsPC LNPs 60 -75 mol%compound 01-1 resulted in the highest protein expression level.
  • Example 17 Screen of %PChemsPC for optimal protein expression
  • PChemsPC LNPs tested 5-10 mol%PChemsPC had the highest protein expression level.
  • the optimal amount of PChemsPC varied with the amount of compound 01-1.
  • the mol%of PChemsPC exceeded 15%, protein expression appeared to decrease.
  • Example 18 Screen of LNP compositions with a steroid containing phospholipid for optimal protein expression
  • Example 19 Tissue-specific Expression of Nucleic Acid Molecules Delivered in LNP formulations.
  • LNP formulations listed in Table 10 containing mRNA encoding luciferase were prepared as described in Example 15.
  • mice Each formulation was systematically administered to 6-8 week old female ICR mice (Xipuer-Bikai, Shanghai) at a 0.25 mg/kg dose by tail vein injection. After 6 hours, the mice were subcutaneously administered with XenoLight D-luciferin (potassium salt) , a substrate ofluciferase that catalyze the production of luminescence. The mice were subsequently euthanized by CO 2 overdoses 15 min thereafter. Mice tissues were harvested and placed in a luminescence imaging scanner to measure the expression level of luciferase in each tissue. The luminescence levels measured from harvest liver tissues were plotted in FIG. 4, showing the mean value and standard deviation (SD) of at least five repeated tested animals for each group.
  • SD standard deviation
  • LNPs composed of a steroid containing phospholipid e.g. PChemsPC, OChemsPC, DChemsPC
  • PChemsPC steroid containing phospholipid
  • DSPC LNP leads to 96%liver distribution with around 3%spleen distribution, while steroid containing phospholipid LNP shows 98 -99%liver distribution, with 0.4%spleen distribution. It can be seen that steroid-modified phospholipid LNP reveals better liver-tropism.
  • Example 20 Characterization of sterol-modified phospholipid LNP with different ionizable lipids
  • LNP formulations containing PChemsPC were prepared with different ionizable lipids.
  • the LNP formulations were composed of a ionizable lipid at a molar ratio of 65%, a PChemsPC lipid at a molar ratio of 10%, a cholesterol-based lipid at a molar ratio of 23.5%and a PEGylated lipid at a molar ratio of 1.5%.
  • Lipid nanoparticles containing human erythropoietin (hEPO) mRNA were prepared as described in Example 15.
  • the effect of replacement of DSPC with PChemsPC on in-vivo protein expression level was also studied.
  • the LNP formulations were composed of an ionizable lipid at a molar ratio of 50%-65%, a phospholipid at a molar ratio of 10%, a cholesterol-based lipid at a molar ratio of 23.5%-38.5%and a PEGylated lipid at a molar ratio of 1.5%.
  • LNP characterizations were listed in Table 12.
  • Example 21 Characterization of in vivo serum cytokines post injection of a steroid containing phospholipid LNP
  • LNP injection will lead to significant increase of pro-inflammatory cytokines, such as inteleukin-6 (IL-6) , tumor necrosis factor-alpha (TNF- ⁇ ) , interferon-gamma (INF- ⁇ ) , interferon-alpha (IFN- ⁇ ) , which can cause innate immune response and leads to undesirable side effects.
  • IL-6 inteleukin-6
  • TNF- ⁇ tumor necrosis factor-alpha
  • IFN- ⁇ interferon-gamma
  • IFN- ⁇ interferon-alpha
  • Lipid nanoparticles containing human erythropoietin (hEPO) mRNA were prepared as described in Example 15, and systemically administered to 6-8 week old female ICR mice (Xipuer-Bikai, Shanghai) at 0.5mg/kg dose by tail vein injection. Mice were euthanized by CO 2 overdoses at 6 hours post administration, and blood samples were taken for cytokines measurement. Particularly, sera were separated from total blood by centrifugation at 5000g for 10 minutes at 4 °C, snap-frozen and stored at -80 °C for analysis.
  • hEPO human erythropoietin
  • DSPC LNPs were composed of ionizable lipid with a molar ratio of 50%, DSPC with a molar ratio of 10%, cholesterol with a molar ratio of 38.5%and a molar ratio of 1.5%for PEGylated lipid.
  • PChemsPC LNPs were composed of ionizable lipid with a molar ratio of 65%, PChemsPC with a molar ratio of 10%, a molar ratio of 23.5%and 1.5%for cholesterol and PEGylated lipids respectively.
  • ionizable lipids e.g. compound 01-1, lipid 5, SM-102, ALC-0315, Compound 03-135) were tested in this study.
  • Example 22 Characterization of in vivo serum cytokines after administration of a steroid containing phospholipid LNP with self-amplifying mRNA (saRNA)
  • lipid nanoparticles containing human erythropoietin (hEPO) self-amplifying mRNA were prepared as described in Example 15. After tail vein injection, mice were euthanized by CO 2 overdoses at 6 hours post administration, and blood samples were taken for cytokines measurement. Cytokine levels were measured and plotted in FIG. 12 and FIG. 13.
  • PChemsPC LNPs lead to significantly lower IL-6, IFN- ⁇ , TNF- ⁇ levels.
  • Example 23 Characterization of sterol-modified phospholipid LNP with CD3-CD19 mRNA
  • DSPC LNPs were composed of compound 01-1 with a molar ratio of 50%, DSPC with a molar ratio of 10%, cholesterol with a molar ratio of 38.5%and a molar ratio of 1.5%for PEGylated lipid.
  • PChemsPC LNPs were composed of compound 01-1 with a molar ratio of 65%, PChemsPC with a molar ratio of 10%, a molar ratio of 23.5%and 1.5%for cholesterol and PEGylated lipids respectively.
  • Lipid nanoparticles encapsulating CD19-CD3 mRNA were prepared as described in Example 15, and systemically administered to 6-8 week old female Balb/c mice (Xipuer-Bikai, Shanghai) at 0.3mg/kg dose by tail vein injection. Mice were euthanized by CO 2 overdoses at 6 hours post administration, and blood samples were taken for antibody measurement. Particularly, serum was separated from total blood by centrifugation at 5000g for 10 minutes at 4 °C, snap-frozen and stored at -80 °C for analysis. The serum antibody level was shown in FIG. 14.
  • PChemsPC LNP can significantly improve CD3-CD19 antibody expression level post administration.

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Abstract

L'invention concerne des LNP contenant des phospholipides comportant une fraction stérol. Les LNP contenant de tels phospholipides ont de potentielles applications dans la technologie des vaccins à ARNm. L'invention concerne des compositions comprenant les LNP et des méthodes d'utilisation des LNP ou des compositions décrites ci-dessus.
EP23905973.6A 2022-12-21 2023-12-20 Nanoparticules lipidiques comprenant des phospholipides modifiés par stérol Pending EP4637718A1 (fr)

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CN2022140569 2022-12-21
PCT/CN2023/140052 WO2024131810A1 (fr) 2022-12-21 2023-12-20 Nanoparticules lipidiques comprenant des phospholipides modifiés par stérol

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WO2022112855A1 (fr) * 2020-11-27 2022-06-02 Guangzhou Ribobio Co., Ltd Composé lipidique et sa composition
WO2022152109A2 (fr) * 2021-01-14 2022-07-21 Suzhou Abogen Biosciences Co., Ltd. Composés lipidiques et compositions de nanoparticules lipidiques
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