WO2024094098A1 - Compound for delivery system and use thereof - Google Patents

Compound for delivery system and use thereof Download PDF

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Publication number
WO2024094098A1
WO2024094098A1 PCT/CN2023/129240 CN2023129240W WO2024094098A1 WO 2024094098 A1 WO2024094098 A1 WO 2024094098A1 CN 2023129240 W CN2023129240 W CN 2023129240W WO 2024094098 A1 WO2024094098 A1 WO 2024094098A1
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group
compound
independently
composition
compound according
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PCT/CN2023/129240
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French (fr)
Chinese (zh)
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夏宇飞
黄箫喃
马艺珊
刘钧健
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益杰立科(上海)生物科技有限公司
中国科学院过程工程研究所
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Publication of WO2024094098A1 publication Critical patent/WO2024094098A1/en

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    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K47/00—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient
    • A61K47/06—Organic compounds, e.g. natural or synthetic hydrocarbons, polyolefins, mineral oil, petrolatum or ozokerite
    • A61K47/16—Organic compounds, e.g. natural or synthetic hydrocarbons, polyolefins, mineral oil, petrolatum or ozokerite containing nitrogen, e.g. nitro-, nitroso-, azo-compounds, nitriles, cyanates
    • A61K47/18—Amines; Amides; Ureas; Quaternary ammonium compounds; Amino acids; Oligopeptides having up to five amino acids
    • 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—Liposomes
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P43/00—Drugs for specific purposes, not provided for in groups A61P1/00-A61P41/00
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07C—ACYCLIC OR CARBOCYCLIC COMPOUNDS
    • C07C275/00—Derivatives of urea, i.e. compounds containing any of the groups, the nitrogen atoms not being part of nitro or nitroso groups
    • C07C275/04—Derivatives of urea, i.e. compounds containing any of the groups, the nitrogen atoms not being part of nitro or nitroso groups having nitrogen atoms of urea groups bound to acyclic carbon atoms
    • C07C275/06—Derivatives of urea, i.e. compounds containing any of the groups, the nitrogen atoms not being part of nitro or nitroso groups having nitrogen atoms of urea groups bound to acyclic carbon atoms of an acyclic and saturated carbon skeleton
    • C07C275/14—Derivatives of urea, i.e. compounds containing any of the groups, the nitrogen atoms not being part of nitro or nitroso groups having nitrogen atoms of urea groups bound to acyclic carbon atoms of an acyclic and saturated carbon skeleton being further substituted by nitrogen atoms not being part of nitro or nitroso groups
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07C—ACYCLIC OR CARBOCYCLIC COMPOUNDS
    • C07C275/00—Derivatives of urea, i.e. compounds containing any of the groups, the nitrogen atoms not being part of nitro or nitroso groups
    • C07C275/04—Derivatives of urea, i.e. compounds containing any of the groups, the nitrogen atoms not being part of nitro or nitroso groups having nitrogen atoms of urea groups bound to acyclic carbon atoms
    • C07C275/18—Derivatives of urea, i.e. compounds containing any of the groups, the nitrogen atoms not being part of nitro or nitroso groups having nitrogen atoms of urea groups bound to acyclic carbon atoms of a saturated carbon skeleton containing rings
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D295/00—Heterocyclic compounds containing polymethylene-imine rings with at least five ring members, 3-azabicyclo [3.2.2] nonane, piperazine, morpholine or thiomorpholine rings, having only hydrogen atoms directly attached to the ring carbon atoms
    • C07D295/02—Heterocyclic compounds containing polymethylene-imine rings with at least five ring members, 3-azabicyclo [3.2.2] nonane, piperazine, morpholine or thiomorpholine rings, having only hydrogen atoms directly attached to the ring carbon atoms containing only hydrogen and carbon atoms in addition to the ring hetero elements
    • C07D295/027—Heterocyclic compounds containing polymethylene-imine rings with at least five ring members, 3-azabicyclo [3.2.2] nonane, piperazine, morpholine or thiomorpholine rings, having only hydrogen atoms directly attached to the ring carbon atoms containing only hydrogen and carbon atoms in addition to the ring hetero elements containing only one hetero ring
    • C07D295/03—Heterocyclic compounds containing polymethylene-imine rings with at least five ring members, 3-azabicyclo [3.2.2] nonane, piperazine, morpholine or thiomorpholine rings, having only hydrogen atoms directly attached to the ring carbon atoms containing only hydrogen and carbon atoms in addition to the ring hetero elements containing only one hetero ring with the ring nitrogen atoms directly attached to acyclic carbon atoms
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D295/00—Heterocyclic compounds containing polymethylene-imine rings with at least five ring members, 3-azabicyclo [3.2.2] nonane, piperazine, morpholine or thiomorpholine rings, having only hydrogen atoms directly attached to the ring carbon atoms
    • C07D295/04—Heterocyclic compounds containing polymethylene-imine rings with at least five ring members, 3-azabicyclo [3.2.2] nonane, piperazine, morpholine or thiomorpholine rings, having only hydrogen atoms directly attached to the ring carbon atoms with substituted hydrocarbon radicals attached to ring nitrogen atoms
    • C07D295/08—Heterocyclic compounds containing polymethylene-imine rings with at least five ring members, 3-azabicyclo [3.2.2] nonane, piperazine, morpholine or thiomorpholine rings, having only hydrogen atoms directly attached to the ring carbon atoms with substituted hydrocarbon radicals attached to ring nitrogen atoms substituted by singly bound oxygen or sulfur atoms

Definitions

  • the present application relates to the field of biomedicine, and specifically to a compound for a delivery system and its use.
  • Therapeutic nucleic acids include small interfering RNA (siRNA), messenger RNA (mRNA), antisense oligonucleotides, ribozymes, DNA enzymes, plasmids, immunostimulatory nucleic acids, antagomirs, antimiRs, mRNA mimics, supermir.UI adapters and aptamers.
  • siRNA small interfering RNA
  • mRNA messenger RNA
  • antisense oligonucleotides include DNA enzymes, plasmids, immunostimulatory nucleic acids, antagomirs, antimiRs, mRNA mimics, supermir.UI adapters and aptamers.
  • Their expression in specific cells is crucial in the treatment of diseases such as those related to protein or enzyme deficiency. Therefore, the effective delivery of these therapeutic nucleic acids to target sites in cells or organisms is a key challenge to achieve nucleic acid-based therapy.
  • oligonucleotide constructs in the current therapeutic environment mainly faces two problems. First, the instability of the structure of free RNA itself makes it extremely easy to be rapidly degraded by various enzymes in the body during delivery; second, the ability of free RNA to enter the intracellular compartment where the relevant translation system is located is limited.
  • Lipid nanoparticles (LNPs) co-formulated with lipids and other lipid components e.g., neutral lipids, steroids, PEGylated lipids
  • LNPs Lipid nanoparticles co-formulated with lipids and other lipid components
  • lipids and other lipid components e.g., neutral lipids, steroids, PEGylated lipids
  • lipids and other lipid components e.g., neutral lipids, steroids, PEGylated lipids
  • the present application provides a compound for a delivery system, including a stereoisomer, a pharmaceutically acceptable salt, a prodrug or a tautomer thereof; and its use, including using the compound alone, or using it in combination with other lipid components such as neutral lipids, charged lipids, steroids and/or their analogs, and/or polymer-conjugated lipids to form lipid nanoparticles for use For delivering therapeutic agents.
  • the compounds of the present application are designed with a substituent group (e.g., urea group) that can form an intermolecular hydrogen bond with the RNA base at the position of 2-6 atoms near the head group (e.g., tertiary amine) that can be protonated in response to changes in pH.
  • a substituent group e.g., urea group
  • the weak interaction force between the nucleic acid formed by this design and the electrostatic effect synergistically load RNA, which can improve the encapsulation efficiency (Encapsulation efficiency) of LNP or liposomes (liposomes), while improving the integrity (Lipid and cargo integrity) of lipids and loaded drugs.
  • the ionizable lipids with urea groups are more conducive to accelerating the degradation of LNP in vivo, thereby improving the safety of the formulation containing the LNP, because the electrophilicity of the hydroxyl group is neutralized by the amines on both sides and does not form an intramolecular six-membered ring or five-membered ring with the tertiary amine.
  • the present application provides a compound, or a pharmaceutically acceptable salt thereof, comprising a cation providing group and a hydrogen bond providing group, and the cation providing group and the hydrogen bond providing group contain 2 to 6 atoms.
  • the compound comprises a cationic lipid.
  • the compound comprises an ionizable lipid.
  • the cation providing group comprises a group that is positively charged at a pH of about 7.0 or less.
  • the cation providing group comprises a group that is positively charged at a pH of about 4.0 or less.
  • the cation-providing group comprises a tertiary amine group.
  • the hydrogen bond providing group comprises a urea group.
  • the compound comprises a tail that is hydrophobic.
  • the tail comprises a saturated and/or unsaturated fatty chain.
  • the tail is substituted on one or both nitrogen atoms of the urea group.
  • the backbone chain length of the tail is 1 to 20 atoms.
  • the compound comprises the following structure (I):
  • L1 is a hydrogen bond providing group
  • G1 is a linking group with a chain length of 2 to 6 atoms
  • said L1 is optionally substituted
  • said G1 is optionally substituted.
  • the compound comprises the following structure (II):
  • G1 is a linking group having a chain length of 2 to 6 atoms, said G1 is optionally substituted, and R1 , R2a and R2b are each independently selected from an optionally substituted substituent.
  • the compound comprises the following structure (IIIa):
  • G1 and G2 are each independently a linking group having a chain length of 2 to 6 atoms, said G1 and G2 are each independently optionally substituted, and R1 , R2a , R2b , R3 , R4a and R4b are each independently selected from optionally substituted substituents.
  • the compound comprises the following structure (IIIb):
  • the G1 is a linking group with a chain length of 2 to 6 atoms, the G1 is optionally substituted, and R1 , R2a , R2b , R5 and R6 are each independently selected from optionally substituted substituents.
  • the compound comprises the following structure (IIIc):
  • G1 and G2 are each independently a linking group having a chain length of 2 to 6 atoms, said G1 and G2 are each independently optionally substituted, and R1 , R2a , R2b , R7 , R8a , R8b and R9 are each independently selected from optionally substituted substituents.
  • the compound comprises the following structure (IIId):
  • G1 , G2 , G3 and G4 are each independently a linking group having a chain length of 2 to 6 atoms, said G1 , G2 , G3 and G4 are each independently optionally substituted, said H1 and H2 are each independently optionally substituted, and R1 , R2a , R2b , R7 , R8a , R8b , R10 , R11a , R11b , R12 , R13a and R13b are each independently selected from optionally substituted substituents.
  • G 1 , G 2 , G 3 and G 4 are each independently a hydrocarbon group, an alicyclic group, and/or an alicyclic heterocyclic group.
  • G 1 , G 2 , G 3 and G 4 are each independently alkyl, alkenyl, and/or alkynyl.
  • one atom in G 1 , G 2 , G 3 and G 4 is each independently replaced by N, O or S.
  • H 1 and H 2 are each independently a hydrocarbon group, an alicyclic group, and/or an alicyclic heterocyclic group.
  • H 1 and H 2 are each independently alkyl, alkenyl, and/or alkynyl.
  • one atom in H1 and H2 is each independently replaced by N, O or S.
  • R 1 , R 2a , R 2b , R 3 , R 4a , R 4b , R 5 , R 6 , R 7 , R 8a , R 8b , R 9 , R 10 , R 11a , R 11b , R 12 , R 13a , and R 13b are each independently a hydrocarbon group, an ester group, an amide group, an N′-alkyl-N-alkenyl-urea group, an alicyclic group, an alicyclic heterocyclic group, an aryl group, and/or a heteroaryl group.
  • R 1 , R 2a , R 2b , R 3 , R 4a , R 4b , R 5 , R 6 , R 7 , R 8a , R 8b , R 9 , R 10 , R 11a , R 11b , R 12 , R 13a , and R 13b are each independently C 1 -C 20 alkyl, C 2 -C 40 alkenyl, C 2 -C 45 ester and/or C 2 -C 20 alkynyl.
  • R 1 , R 2a , R 2b , R 3 , R 4a , R 4b , R 5 , R 6 , R 7 , R 8a , R 8b , R 9 , R 10 , R 11a , R 11b , R 12 , R 13a , and R 13b are each independently branched or unbranched.
  • R 1 , R 2a , R 2b , R 3 , R 4a , R 4b , R 5 , R 6 , R 7 , R 8a , R 8b , R 9 , R 10 , R 11a , R 11b , R 12 , R 13a , and R 13b each independently contain 0 to 4 unsaturated bonds.
  • R 1 , R 2a , R 2b , R 3 , R 4a , R 4b , R 5 , R 6 , R 7 , R 8a , R 8b , R 9 , R 10 , R 11a , R 11b , R 12 , R 13a , and R 13b are each independently substituted by R x , and R x is an optionally substituted substituent.
  • R x is C 1 -C 20 alkyl, C 2 -C 25 ester, C 2 -C 20 alkenyl, amide, urea, hydroxyl, sulfhydryl or amino.
  • the present application provides a compound, or a pharmaceutically acceptable salt thereof, wherein the compound is selected from the structures provided in the present application.
  • the present application provides a composition comprising the compound and a therapeutic agent.
  • the composition further comprises a substance selected from the group consisting of a neutral lipid, a steroid, and a polymer-conjugated lipid.
  • the composition comprises liposomes.
  • the composition is about 500 nm or less in diameter.
  • the steroid comprises cholesterol.
  • the polymer-conjugated lipid comprises a PEGylated lipid.
  • the molar ratio of the compound to the neutral lipid is from about 2:1 to about 8:1.
  • the molar ratio of the compound to the steroid is about 5:1 to 1:1.
  • the molar ratio of the compound to the polymer-conjugated lipid is about 100:1 to about 20:1.
  • the therapeutic agent comprises a nucleic acid.
  • the therapeutic agent comprises a component of a CRISPR system and/or a nucleic acid encoding a component of a CRISPR system.
  • the therapeutic agent comprises a guide RNA and/or a nucleic acid encoding a Cas enzyme.
  • the present application provides a cell comprising the compound and/or the composition.
  • the present application provides a kit comprising the compound, the composition and/or the cell.
  • the present application provides a method for administering a therapeutic agent, which comprises providing the compound, the composition, the cell and/or the kit, wherein the therapeutic agent is mixed with the compound, or the therapeutic agent is present in the composition, the cell and/or the kit.
  • the present application provides a method for delivering nucleic acid, which comprises providing the compound, the composition, the cell and/or the kit, wherein the nucleic acid is mixed with the compound, or the nucleic acid is present in the composition, the cell and/or the kit.
  • the present application provides a use of the compound, the composition, the cell and/or the kit in the preparation of a drug, wherein the drug is used in a method for treating and/or preventing a disease or condition.
  • FIG. 1 shows the expression effect of the delivery substance of the present application on Luciferase in mouse liver.
  • FIG2 shows the evaluation of the effect of intravenous injection of the delivery material of the present application on liver transfection in adult Ai9 transgenic mice.
  • FIG3 shows the evaluation of the PCSK9 gene editing effect of the delivered material of the present application in vivo in mice.
  • the term "compound” generally refers to a substance having two or more different elements.
  • the compound of the present application may be an organic compound, for example, the compound of the present application may be a compound with a molecular weight of less than 500, a compound with a molecular weight of less than 1000, a compound with a molecular weight of more than 1000, or a compound with a molecular weight of more than 10000 or more than 100000.
  • a compound may also refer to a compound connected by chemical bonds, for example, a compound in which one or more molecules with a molecular weight of less than 1000 are connected to a biomacromolecule by chemical bonds, and the biomacromolecule may be a high polysaccharide, protein, nucleic acid, polypeptide, etc.
  • the compound of the present application may include a compound in which a protein is connected to one or more molecules with a molecular weight of less than 1000, a compound in which a protein is connected to one or more molecules with a molecular weight of less than 10000, or a compound in which a protein is connected to one or more molecules with a molecular weight of less than 100000.
  • the term "compound of the present application” refers to the compound of the present application.
  • the term also includes various crystalline forms, pharmaceutically acceptable salts, hydrates or solvates of the compound of the present application.
  • prodrug generally refers to the prodrug substance that can be metabolized to form a substance having the structure of the compound of the present application after being administered to a subject
  • metabolite generally refers to the substance obtained by the metabolism of the compound of the present application after being administered to a subject, and the prodrugs and metabolites contained in such derivatives are included within the scope of the present invention.
  • the compounds of the present application include tautomers, mesomorphs, racemates, enantiomers, and/or diastereomers of the compounds.
  • the term “diastereomer” generally refers to a stereoisomer having two or more chiral centers and whose molecules are not mirror images of each other. Diastereomers can have different physical properties, such as melting points, boiling points, spectral properties, and reactivity.
  • the terms "tautomer” or “tautomeric form” are used interchangeably and generally refer to structural isomers of different energies that can be mutually converted by low energy barriers.
  • proton tautomers include mutual conversions by proton migration, such as keto-enol isomerization and imine-enamine isomerization.
  • Valence tautomers include mutual conversions by the reorganization of some bonding electrons.
  • the term “mesomeric compound” generally refers to a molecule containing an asymmetric atom, but due to its symmetry factors, the total optical rotation of the molecule is zero.
  • the term “racemic compound” or “racemic mixture” refers to a composition composed of equimolar amounts of two enantiomeric substances.
  • each chiral carbon atom can optionally be in R configuration or S configuration, or a mixture of R configuration and S configuration.
  • some atoms of the compounds of the present application may appear in more than one isotopic form.
  • hydrogen may exist in the form of protium ( 1H ), deuterium ( 2H ) and tritium ( 3H ), and carbon may exist naturally in three different isotopes ( 12C , 13C and 14C ).
  • isotopes that can be incorporated into the compounds of the present application also include but are not limited to15N , 18O , 17O , 18F , 32P , 33P , 129I , 131I , 123I , 124I , 125I , or similar isotopes.
  • the compounds of the present application can be enriched in one or more of these isotopes.
  • such isotope-enriched compounds can be used for a variety of purposes.
  • substitution with heavy isotopes such as deuterium ( 2H ) may provide certain therapeutic advantages, which may be due to higher metabolic stability.
  • the natural abundance of deuterium ( 2H ) is about 0.015%. Therefore, there is one deuterium atom for approximately every 6500 hydrogen atoms in nature. Therefore, the deuterium-containing compounds of the present application have a deuterium abundance greater than 0.015% at one or more positions (as the case may be).
  • the structures described in the present application may also include compounds that differ only in the presence or absence of one or more isotopically enriched atoms.
  • compounds that are consistent with the structures of the present application except that hydrogen atoms are replaced by deuterium or tritium, or carbon atoms are replaced by carbon 13 or carbon 14, are all in Within the scope of this application.
  • the term “pharmaceutically acceptable” refers to substances that are suitable for contact with human and animal tissues without excessive toxicity, irritation, allergic reaction or other problems or complications, and are commensurate with a reasonable benefit/risk ratio, within the scope of reasonable medical judgment.
  • pharmaceutically acceptable salt refers to a salt of the compound that is pharmaceutically acceptable as defined above and has the desired pharmacological activity, including acid addition salts and base addition salts of the compound.
  • pharmaceutically acceptable acid addition salt refers to a free base which retains the biological effectiveness and properties of the free base, is not biologically or otherwise undesirable, and is soluble in water and is soluble in water and is stable to water and is stable to water.
  • the salts formed by the pharmaceutically acceptable base addition salts include maleic acid, galactaric acid, gentisic acid, glucoheptonic acid, glucuronic acid, glutamic acid, glutaric acid, 2-oxo-glutaric acid, glycerophosphoric acid, glycolic acid, hippuric acid, isobutyric acid, lactic acid, lactobionic acid, lauric acid, maleic acid, malic acid, malonic acid, mandelic acid, methanesulfonic acid, mucic acid, naphthalene-1,5-disulfonic acid, naphthalene-2-sulfonic acid, 1-hydroxy-2-naphthoic acid, nicotinic acid, oleic acid, orotic acid, oxalic acid, palmitic acid, pamoic acid, propionic acid, pyroglutamic acid, pyruvic acid, salicylic acid, 4-aminosalicylic acid, sebacic acid, stearic
  • salts derived from inorganic bases include, but are not limited to, sodium, potassium, lithium, ammonium, calcium, magnesium, iron, zinc, copper, manganese, aluminum, etc.
  • Salts derived from organic bases include, but are not limited to, salts of primary, secondary, and tertiary amines, substituted amines including naturally occurring substituted amines, cyclic amines, and basic ion exchange resins such as ammonia, isopropylamine, trimethylamine, diethylamine, triethylamine, tripropylamine, diethanolamine, ethanolamine, diarnol, 2-dimethylaminoethanol, 2-diethylaminoethanol, dicyclohexylamine, lysine, arginine, histidine, caffeine, procaine, hydrazine, choline, betaine, benzylidene, benzathine, ethylenediamine, glucosamine, methylglucamine, theobromine, triethanolamine, tromethamine, purine, piperazine, piperidine, N-ethylpiperidine, polyamine resins, and the like.
  • groups containing 2 to 6 atoms and “linking groups with a chain length of 2 to 6 atoms” refer to 2, 3, 4, 5 or 6 atoms connected to each other via continuous covalent bonds to form a chain-like molecular fragment or linking group, wherein the atoms may be single atoms (such as but not limited to carbon atoms, nitrogen atoms, oxygen atoms and sulfur atoms, etc.), central atoms contained in any substituent (such as but not limited to the central carbon atom of an alkyl, alkenyl or alkynyl group and the central nitrogen atom of an amine group, etc.) and/or atoms participating in the formation of the main chain in any substituent (such as but not limited to the carbon atoms and oxygen atoms connected by single bonds in an ester group, etc.).
  • substituent such as but not limited to the central carbon atom of an alkyl, alkenyl or alkynyl group and the central nitrogen atom of an amine group, etc.
  • the groups may contain multiple atoms forming a ring structure or the linking group between the groups is a ring group, in which case the number of atoms or the chain length of the spacer is the number of atoms between the two atoms connected to the main chain on the ring structure. The number of atoms that form the shortest distance between the atoms (including the atoms at both ends).
  • the term "ionizable lipid” refers to a lipid having at least one protonatable or deprotonatable group, such as a cationic lipid, such that the lipid is positively charged at a pH equal to or lower than physiological pH (e.g., pH 7.4) and is neutral at a second pH (preferably equal to or higher than physiological pH).
  • physiological pH e.g., pH 7.4
  • second pH preferably equal to or higher than physiological pH
  • the pKa of the protonatable group of an ionizable lipid is in the range of about 4 to about 7, and at different pHs, its ionization reaction affects the surface charge of the LNP, and this charge state can affect the absorption of plasma proteins, blood clearance, and tissue distribution, which are critical for intracellular delivery of nucleic acids, as well as the ability to form endosomolytic non-bilayer structures.
  • cation providing group generally refers to a head group with a positive charge in an ionizable lipid, and its group size and charge density have an impact on the efficiency of encapsulating nucleic acids, stabilizing LNP, interacting with cell membranes, and promoting processes such as endosome escape.
  • Common ionizable lipids contain only one head group, and some also contain several head groups.
  • Non-limiting examples of cation providing groups include amines (primary amines, secondary amines, tertiary amines, quaternary amines), guanidines, heterocyclic groups, etc.
  • the ionizable lipids DLin-MC3-DMA, SM-102, and ALC-0315 used clinically contain tertiary amine heads, which can undergo pH-dependent cationization.
  • the cation providing group can be substituted or non-substituted.
  • hydrogen bond providing group generally refers to a linking group or a group as part of a linking group in an ionizable lipid that can form a weak intermolecular interaction force with a base of a nucleic acid as a hydrogen bond donor
  • hydrogen bond providing groups include urea, amide, carboxyl, divalent transition metal, etc.
  • the hydrogen bond providing group may be substituted or unsubstituted.
  • the term "the tail has hydrophobicity" refers to a tail group having a non-polar group
  • the non-polar group includes, but is not limited to, long-chain saturated and unsaturated aliphatic hydrocarbon groups, and such groups optionally substituted with one or more aromatic, cycloaliphatic or heterocyclic groups.
  • Preferred examples include, but are not limited to, diacylglycerol, dialkylglycerol, N-N-dialkylamino, 1,2-diacyloxy-3-aminopropane and 1,2-dialkyl-3-aminopropane.
  • main chain in the case of a certain group generally refers to a carbon chain with more carbon atoms, or the longest carbon chain containing a functional group as the main chain.
  • the carbon chain with the largest number of branches is selected as its main chain.
  • the terms "optionally substituted substituents" and “optionally substituted” mean that the referenced group may be substituted or unsubstituted by one or more additional groups individually and independently selected from the group consisting of alkyl, alkenyl, alkynyl, cycloalkyl, aryl, heteroaryl, heterocycloalkyl, hydroxy, alkoxy, mercapto, cyano, halogen, carbonyl, thiocarbonyl, isocyanato, thiocyanato, isothiocyanato, nitro, perhaloalkyl, perfluoroalkyl and mono- and di-substituted amino groups.
  • the amino group and its protected derivatives The amino group and its protected derivatives.
  • alkyl generally refers to an alkane residue derived from a hydrogen atom. Alkyl can be substituted or non-substituted, substituted or non-substituted.
  • alkyl generally refers to a saturated straight or branched aliphatic hydrocarbon radical, which has a residue derived from a hydrogen atom removed from the same carbon atom or two different carbon atoms of the parent alkane, which can be a straight or branched group containing 1 to 20 carbon atoms, for example, containing 1 to 12 carbon atoms, for example, a chain alkyl containing 1 to 6 carbon atoms.
  • alkyl include, but are not limited to, methyl, ethyl, propyl, propyl, butyl, etc.
  • the alkyl group may be substituted or unsubstituted, substituted or unsubstituted, for example, when substituted, the substituent may be substituted at any available point of attachment, and the substituent may be independently selected from one or more substituents selected from alkyl, alkenyl, alkynyl, alkoxy, alkylthio, alkylamino, halogen, mercapto, hydroxy, nitro, cyano, cycloalkyl, heterocyclyl, aryl, heteroaryl, cycloalkyloxy, heterocycloalkyloxy, cycloalkylthio, heterocycloalkylthio and oxo, for example, hydrogen, protium, deuterium, tritium, halogen, -NO2 , -CN, -OH, -SH, -NH
  • alkenyl generally refers to a straight or branched hydrocarbon group containing one or more double bonds.
  • alkenyl include allyl, homoallyl, vinyl, crotyl, butenyl, pentenyl and hexenyl, etc.
  • C2-6 alkenyl with more than one double bond include butadienyl, pentadienyl, hexadienyl and hexatrienyl and their branched forms.
  • the position of the unsaturated bond (double bond) can be at any position of the carbon chain.
  • Alkenyl can be substituted or unsubstituted.
  • alkynyl generally refers to an unsaturated straight or branched chain alkynyl group, for example, ethynyl, 1-propynyl, propargyl, butynyl, etc.
  • the alkynyl group may be substituted or unsubstituted.
  • hydrocarbyl generally refers to a straight or branched chain or cyclic hydrocarbon radical or combinations thereof consisting only of carbon atoms and hydrogen atoms, which may be fully saturated, monounsaturated or polyunsaturated, and may include divalent and polyvalent radicals, having a specified number of carbon atoms (i.e., C1 - C10 means one to ten carbons).
  • saturated hydrocarbon radicals include, but are not limited to Homologs and isomers of methyl, ethyl, n-propyl, isopropyl, n-butyl, tert-butyl, isobutyl, sec-butyl, cyclohexyl, (cyclohexyl)methyl, cyclopropylmethyl, n-pentyl, n-hexyl, n-heptyl, n-octyl, etc.
  • Unsaturated hydrocarbon groups are those having one or more double bonds or triple bonds.
  • unsaturated hydrocarbon groups include, but are not limited to, vinyl, 2-propenyl, crotyl, 2-isopentenyl, 2-(butadienyl), 2,4-pentadienyl, 3-(1,4-pentadienyl), ethynyl, 1- and 3-propynyl, 3-butynyl, and higher homologs and isomers.
  • hydrocarbon group is also meant to include those hydrocarbon derivatives defined in detail below, such as “heterohydrocarbon groups", unless otherwise noted. Hydrocarbon groups limited to hydrocarbon groups are also referred to as "homohydrocarbon groups”.
  • alicyclic group generally refers to a residue derived from the same carbon atom or multiple different carbon atoms of an aliphatic ring by removing a hydrogen atom.
  • cycloalkane generally refers to a saturated or partially unsaturated monocyclic or polycyclic cyclic hydrocarbon, and the carbocyclic ring contains 3 to 20 carbon atoms, can contain 3 to 12 carbon atoms, can contain 3 to 10 carbon atoms, and can contain 3 to 8 carbon atoms.
  • Non-limiting examples of alicyclic groups include cyclopropane, cyclobutane, cyclopentane, cyclopentenyl, cyclohexane, cyclohexenyl, cyclohexadienyl, cycloheptane, cycloheptatrienyl, cyclooctane, etc.; polycyclic carbocyclic rings can include spirocyclic, condensed and bridged carbocyclic rings. Alicyclic groups can be substituted or unsubstituted.
  • the term "carbocyclic group” generally refers to a residue derived from the removal of a hydrogen atom from a carbon atom of a carbocyclic ring.
  • Carbocycle generally refers to a saturated or partially unsaturated monocyclic or polycyclic cyclic hydrocarbon, the carbocycle containing 3 to 20 carbon atoms, can contain 3 to 12 carbon atoms, can contain 3 to 10 carbon atoms, can contain 3 to 8 carbon atoms.
  • Non-limiting examples of monocyclic carbocycles include cyclopropane, cyclobutane, cyclopentane, cyclopentene, cyclohexane, cyclohexene, cyclohexadiene, cycloheptane, cycloheptatriene, cyclooctane, etc.; polycyclic carbocycles can include spirocyclic, condensed and bridged carbocycles. Carbocyclic groups can be substituted or unsubstituted. In some cases, alicyclic and carbocyclic rings can be used interchangeably.
  • partially unsaturated generally refers to a ring structure containing at least one double bond or triple bond between the ring molecules.
  • the term “partially unsaturated” encompasses ring structures with multiple unsaturations, but is not intended to include aromatic or heteroaromatic rings as defined in this application.
  • the term “unsaturated” means that the moiety has one or more degrees of unsaturation.
  • alicyclic group generally refers to a stable 3-7-membered monocyclic carbon ring structure without aromaticity, a fused 7-10-membered bicyclic heterocyclic structure or a bridged 6-10-membered bicyclic heterocyclic structure, which ring structures can be saturated or partially saturated.
  • these ring structures also contain one or more heteroatoms, wherein the heteroatoms can be selected from the following groups: oxygen, sulfur and nitrogen. For example, it contains 1-4 heteroatoms as defined above.
  • nitrogen can include nitrogen that has undergone substitution reactions.
  • the alicyclic group can include "heterocycloalkyl", which can refer to a stable 3-7-membered monocyclic alkane structure without aromaticity, a fused 7-10-membered bicyclic heterocyclic structure or a bridged 6-10-membered bicyclic heterocyclic structure, which can contain one or more heteroatoms in addition to carbon atoms, wherein the heteroatoms can be selected from the following groups: oxygen, sulfur and nitrogen. For example, it contains 1-4 heteroatoms as defined above. Heterocycloalkyl groups may be substituted or unsubstituted. Alicyclic heterocyclic groups may be substituted or unsubstituted.
  • aryl generally refers to a residue derived from an aromatic ring by removing a hydrogen atom.
  • aromatic ring may refer to a 6 to 14-membered all-carbon monocyclic ring or a fused polycyclic ring (i.e., a ring sharing adjacent carbon atom pairs) with a conjugated ⁇ electron system, which may be 6 to 10 members, such as benzene and naphthalene.
  • the aromatic ring may be fused to a heteroaryl, heterocyclic or cycloalkyl ring, wherein the ring connected to the parent structure is an aryl ring.
  • the aryl group may be substituted or unsubstituted, and when substituted, the substituent may be one or more of the following groups, independently selected from the following groups: alkyl, alkenyl, alkynyl, alkoxy, alkylthio, alkylamino, halogen, sulfhydryl, hydroxyl, nitro, cyano, cycloalkyl, heterocycloalkyl, aryl, heteroaryl, cycloalkyloxy, heterocycloalkyloxy, cycloalkylthio, and heterocycloalkylthio.
  • the aryl group may be substituted or unsubstituted.
  • heteroaryl generally refers to a residue derived from a carbon atom of a heteroaromatic ring by removing a hydrogen atom.
  • heteromatic ring refers to a heteroaromatic system containing 1 to 4 heteroatoms, 5 to 14 ring atoms, wherein the heteroatoms can be selected from the following groups: oxygen, sulfur and nitrogen.
  • the heteroaryl group can be 5 to 10 yuan, and can be 5 yuan or 6 yuan, such as furanyl, thienyl, pyridyl, pyrrolyl, N-alkylpyrrolyl, pyrimidyl, pyrazinyl, imidazolyl, tetrazolyl, etc.
  • the heteroaryl ring can be fused to an aryl, heterocyclyl or cycloalkyl ring, wherein the ring connected to the parent structure is a heteroaryl ring.
  • Heteroaryl can be optionally substituted or unsubstituted, and when substituted, the substituent can be one or more of the following groups, which are independently selected from the following groups: alkyl, alkenyl, alkynyl, alkoxy, alkylthio, alkylamino, halogen, sulfhydryl, hydroxyl, nitro, cyano, cycloalkyl, heterocycloalkyl, aryl, heteroaryl, cycloalkyloxy, heterocycloalkyloxy, cycloalkylthio, and heterocycloalkylthio. Heteroaryl can be substituted or unsubstituted.
  • alkyl alkenyl
  • cycloalkyl alkyl
  • terms such as “alkyl”, “alkenyl”, “cycloalkyl”, etc. may be preceded by an identifier to indicate the number of atoms present in the group in a particular case, for example, C 1 -C 4 alkyl, C 3 -C 7 cycloalkyloxy, C 1 -C 4 alkylcarbonylamino, etc., and the subscript number following "C” indicates the number of carbon atoms present in the group.
  • C 3 alkyl refers to an alkyl group having three carbon atoms (e.g., n-propyl, isopropyl); in C 1-10 , the members of the group may have any number of carbon atoms falling within the range of 1-10.
  • One or more hydrogen atoms in the group for example, up to 5, for example, 1-3 hydrogen atoms are replaced by the corresponding number of substituents independently of each other.
  • the substituents are only in their possible chemical positions, and those skilled in the art can determine (by experiment or theory) possible or impossible replacements without paying too much effort.
  • amino or hydroxyl groups with free hydrogen may be unstable when combined with carbon atoms with unsaturated (such as olefinic) bonds.
  • neutral lipid refers to any of a variety of lipid substances that exist in an uncharged or neutral zwitterionic form at a selected pH.
  • lipids include, but are not limited to, phosphatidylcholines such as 1,2-distearoyl-sn-glycero-3-phosphocholine (DSPC), 1,2-dipalmitoyl-sn-glycero-3-phosphocholine (DPPC), 1,2-dimyristoyl-sn-glycero-3-phosphocholine (DMPC), 1-palmitoyl-2-oleoyl-sn-glycero-3-phosphocholine (POPC), 1,2-dioleoyl-sn-glycero-3-phosphocholine (DOPC), phosphatidylethanolamines such as 1,2-dioleoyl-sn- Glyceryl-3-phosphoethanolamine (DOPE), sphingomyelin (SM), ceramides
  • DOPE 1,2-dioleoy
  • Non-limiting examples thereof include cholesterol and the like.
  • polymer-conjugated lipid refers to a molecule comprising a lipid portion and a polymer portion.
  • An example of a polymer-conjugated lipid is a pegylated lipid.
  • pegylated lipid refers to a molecule comprising a lipid portion and a polyethylene glycol portion. Pegylated lipids are known in the art and include 1-(monomethoxy-polyethylene glycol)-2,3-dimyristoylglycerol (PEG-DMG) and the like.
  • the term “liposome” generally refers to a vesicle with an internal space that is isolated from the external medium by one or more bilayer, multilayer, any lamellar or solid membranes.
  • the bilayer membrane can be formed by amphiphilic molecules, such as synthetic or natural lipids containing spatially isolated hydrophilic and hydrophobic domains; for another example, the bilayer membrane can be formed by amphiphilic polymers and surfactants.
  • Liposomes encapsulate an aqueous phase, which generally contains substances to be delivered to cells, such as nucleic acids.
  • the term "liposome” as used in the present application also covers lipid- and polymer-based nanoparticles.
  • nucleic acid refers to a polymer containing at least two deoxyribonucleotides or ribonucleotides in a single-stranded or double-stranded form, and includes DNA, RNA and hybrids thereof.
  • DNA can be in the form of antisense molecules, plasmid DNA, cDNA, PCR products or vectors.
  • RNA can be in the form of small hairpin RNA (shRNA), messenger RNA (mRNA), antisense RNA, miRNA, micRNA, multivalent RNA, Dicer substrate RNA or viral RNA (vRNA) and combinations thereof.
  • Nucleic acid includes nucleic acids containing known nucleotide analogs or modified main chain residues or bonds, which are synthetic, naturally occurring and non-naturally occurring and have binding properties similar to reference nucleic acids.
  • Examples of such analogs include, but are not limited to, phosphorothioates, phosphoramidates, methyl phosphonates, chiral-methyl phosphonates, 2'-0-methyl ribonucleotides and peptide-nucleic acids (PNA).
  • PNA peptide-nucleic acids
  • the term encompasses nucleic acids containing known analogs of natural nucleotides that have similar binding properties to reference nucleic acids.
  • nucleic acid sequence also implicitly encompasses conservatively modified variants thereof (e.g., degenerate codon substitutions), alleles, orthologs, single nucleotide polymorphisms, and complementary sequences as well as the sequence explicitly indicated.
  • a "nucleotide” contains the following sugars: deoxyribose (DNA) or ribose (RNA); a base and a phosphate group. The nucleotides are linked together by the phosphate groups.
  • Bases include purines and pyrimidines, which further include the natural compounds adenine, thymine, guanine, cytosine, uracil, inosine, and natural analogs, as well as synthetic derivatives of purines and pyrimidines, Modifications include, but are not limited to, placement of new reactive groups such as, but not limited to, amines, alcohols, thiols, carboxylates, and hydrocarbyl halides.
  • administering refers to introducing a composition or medicament (e.g., nucleic acid) into an individual and includes introducing one or more compositions or medicaments simultaneously and sequentially.
  • administering may refer to, for example, treatment, pharmacokinetics, diagnosis, research, placebo, and experimental methods.
  • administering also encompasses in vitro and ex vivo treatments.
  • Introducing a composition or medicament into an individual is by any suitable route, including oral, pulmonary, nasal, parenteral (intravenous, intramuscular, intraperitoneal or subcutaneous), rectal, intralymphatic, intratumoral or local.
  • Administration includes self-administration and administration by another person. Administration may be performed by any suitable route. Suitable administration routes allow a composition or medicament to perform its intended function. For example, if the suitable route is intravenous, then administering the composition by introducing the composition or medicament into the vein of an individual.
  • delivering a therapeutic and/or prophylactic agent to a subject may involve administering a nanoparticle composition comprising a therapeutic and/or prophylactic agent to the subject (e.g., by intravenous, intramuscular, intradermal or subcutaneous routes).
  • Administering a nanoparticle composition to a mammal or mammalian cell may involve contacting one or more cells with the nanoparticle composition.
  • contact refers to establishing a physical connection between two or more entities, and methods for contacting a cell with an external entity in vivo and in vitro are well known in the biological field.
  • the term "prevent” or “preventing” encompasses the following situations: (1) inhibiting the onset of a disease in a subject or patient who may be at risk for and/or susceptible to the disease but who does not yet experience or display any or all of the symptoms or signs of the disease; and/or (2) slowing the onset of symptoms or signs of a disease in a subject or patient who may be at risk for and/or susceptible to the disease but who does not yet experience or display any or all of the symptoms or signs of the disease.
  • treating encompasses: (1) inhibiting a disease in a subject or patient who is experiencing or exhibiting symptoms or signs of the disease (e.g., arresting further development of the symptoms and/or signs), (2) ameliorating a disease in a subject or patient who is experiencing or exhibiting symptoms or signs of the disease (e.g., reversing the symptoms and/or signs), and/or (3) achieving any measurable reduction in a disease in a subject or patient who is experiencing or exhibiting symptoms or signs of the disease.
  • the term "subject” or “patient” refers to any organism to which the compounds and/or compositions described herein can be administered, for example, for experimental, diagnostic, preventive and/or therapeutic purposes, and typical subjects include animals (e.g., mammals such as mice, rats, rabbits, non-human primates and humans) and/or plants.
  • animals e.g., mammals such as mice, rats, rabbits, non-human primates and humans
  • the term "about” generally refers to a variation within the range of 0.5%-10% above or below the specified value, for example, 0.5%, 1%, 1.5%, 2%, 2.5%, 3%, 3.5%, 4%, 4.5%, 5%, 5.5%, 6%, 7%, 8%, 9%, 10%, 11%, 12%, 13%, 14%, 15%, 16%, 17%, 18%, 19%, 20%, 21%, 22%, 23%, 24%, 25%, 26%, 27%, 28%, 29%, 30%, 31%, 32%, 33%, 34%, 35%, 36%, 37%, 38%, 39%, 40%, 41%, 42%, 43%, 44%, 45%, 46%, 47%, 48%, 49%, 50%, 51%, 52%, 53%, 54%, 55%, 56%, 57%, 58%, 59%, 60%, 61%, 62%, 63%, 64%, 65%, 66%, 67%, 68%, 69%, 70%, 71%
  • the present application provides a compound, or a pharmaceutically acceptable salt thereof, comprising a cation providing group and a hydrogen bond providing group, wherein 2 to 6 atoms are contained between the cation providing group and the hydrogen bond providing group, for example, 2, 3, 4, 5, or 6 atoms are contained between the cation providing group and the hydrogen bond providing group of the compound.
  • the compound comprises a cationic lipid.
  • the compound comprises an ionizable lipid.
  • the cation providing group includes a group that is positively charged at a pH of about 7.0 or lower.
  • the cation providing group is a group that is positively charged at the following pH conditions: about 7.0, about 6.0, about 5.0, about 4.0, about 3.0, about 2.0, about 1.0, about 0.9, about 0.8, about 0.7, about 0.6, about 0.5, about 0.4, about 0.2, about 0.1.
  • the cation providing group includes a group that is positively charged at a pH of about 4.0 or lower.
  • the cation providing group is a group that is positively charged at the following pH conditions: about 4.0, about 3.0, about 2.0, about 1.0, about 0.9, about 0.8, about 0.7, about 0.6, about 0.5, about 0.4, about 0.3, about 0.2, about 0.1.
  • the cation providing group comprises a tertiary amine group, wherein the three hydrocarbon groups connected to the amine of the trivalent group in the tertiary amine group are independently selected from optionally substituted substituents.
  • the hydrogen bond providing group comprises a urea group, wherein the two hydrocarbon groups (i.e., a total of four hydrocarbon groups) respectively connected to the amines of the two trivalent groups in the urea group are independently selected from optionally substituted substituents.
  • the compound comprises a tail, and the tail is hydrophobic.
  • the tail is substituted on one or two nitrogen atoms of the urea group.
  • the tail comprises a saturated fatty chain.
  • the tail comprises an unsaturated fatty chain.
  • the tail comprises an unsaturated fatty chain, and the unsaturation is partially unsaturated.
  • the tail comprises an unsaturated fatty chain, and the unsaturation is monounsaturated.
  • the tail comprises an unsaturated fatty chain, and the unsaturation is polyunsaturated.
  • the main chain length of the tail is 1 to 20 atoms.
  • the tail has a main chain length of 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, or 20 atoms.
  • the compound comprises structure (I):
  • L1 is the hydrogen bond providing group, and the L1 is optionally substituted
  • G1 is a linking group with a chain length of 2 to 6 atoms, and the G1 is optionally substituted.
  • G1 is a linking group with the following chain lengths: 2, 3, 4, 5, 6 atoms.
  • G1 is selected from any of the following groups or any combination thereof: hydrocarbon group, alicyclic group, alicyclic group, alkyl group, alkenyl group, alkynyl group.
  • G1 is optionally substituted, and one atom is replaced by N, replaced by O, or replaced by S.
  • the structure of the compound is as follows (as described in structure OT13-1 provided herein):
  • the compound comprises any one of structure (II) and structure (IIIb):
  • G1 is a linking group with a chain length of 2 to 6 atoms, and G1 is optionally substituted, and R1 , R2a , R2b , R5 and R6 are each independently optionally substituted.
  • G1 is a linking group with the following chain lengths: 2, 3, 4, 5, 6 atoms.
  • G1 is selected from any of the following groups or any combination thereof: hydrocarbon group, alicyclic group, alicyclic group, alkyl group, alkenyl group, alkynyl group.
  • G1 is optionally substituted, and one atom is replaced by N, replaced by O or replaced by S.
  • R 1 , R 2a , R 2b , R 5 and R 6 are each independently selected from the following groups or any combination thereof: hydrocarbon group, ester group, amide group, N'-alkyl-N-alkenyl-urea group, alicyclic group, alicyclic heterocyclic group, aryl group, heteroaryl group, C 1 alkyl group, C 2 alkyl group, C 3 alkyl group, C 4 alkyl group, C 5 alkyl group, C 6 alkyl group, C 7 alkyl group, C 8 alkyl group, C 9 alkyl group, C 10 alkyl group, C 11 alkyl group, C 12 alkyl group, C 13 alkyl group, C 14 alkyl group, C 15 alkyl group, C 16 alkyl group, C 17 alkyl group, C 18 alkyl group, C 19 alkyl group, C 20 alkyl group, C 2 alkenyl group, C 3 alkenyl group, C 4 alkenyl group, C
  • R1 , R2a , R2b , R5 and R6 are each independently branched or unbranched.
  • R 1 , R 2a , R 2b , R 5 and R 6 each independently contain 0, 1, 2, 3 or 4 unsaturated bonds.
  • R 1 , R 2a , R 2b , R 5 and R 6 are each independently substituted by an optionally substituted substituent RX .
  • R 1 , R 2a , R 2b , R 5 and R 6 are each independently substituted by an optionally substituted substituent RX , and RX is selected from any of the following groups: C 1 -C 20 alkyl, C 2 -C 25 ester, C 2 -C 20 alkenyl, amide, urea, hydroxyl, thiol, amino.
  • the structure of the compound is as follows (as described in structure OT13-3 provided herein):
  • the compound comprises a tertiary amine group as a cation providing group and comprises the structure (II), wherein G1 is a C3 hydrocarbon group with a chain length of 3 atoms, connected to G1 is a hydrogen bond providing group urea group, and the amines of the two trivalent groups of the urea group are independently substituted, that is, R1 in the structure (II) is a branched alkyl ester substituent, and its molecular formula is CH ( C8H16COOCH2C6H13C4H9 ) 2 , and R2a and R2b in the structure (II ) are hydrogen and C8 alkyl respectively (in any order).
  • the compound comprises any one of structure (IIIa) and structure (IIIc):
  • G1 and G2 are each independently a linking group with a chain length of 2 to 6 atoms, and G1 and G2 are each independently optionally substituted, and R1 , R2a , R2b , R3 , R4a , R4b , R7 , R8a , R8b and R9 are each independently optionally substituted.
  • G1 and G2 are each independently a linking group with a chain length of 2, 3, 4, 5, 6 atoms.
  • G1 and G2 are each independently selected from any of the following groups or any combination thereof: hydrocarbon, alicyclic, alicyclic, alkyl, alkenyl, alkynyl.
  • G1 and G2 are each independently optionally substituted, and one atom is replaced by N, replaced by O or replaced by S.
  • R 1 , R 2a , R 2b , R 3 , R 4a , R 4b , R 7 , R 8a , R 8b and R 9 are each independently selected from the following groups or any combination thereof: a hydrocarbon group, an ester group, an amide group, an N′-alkyl-N-alkenyl-urea group, an alicyclic group, an alicyclic group, an aryl group, a heteroaryl group, a C 1 alkyl group, a C 2 alkyl group, a C 3 alkyl group, a C 4 alkyl group, a C 5 alkyl group, a C 6 alkyl group, a C 7 alkyl group, a C 8 alkyl group, a C 9 alkyl group, a C 10 alkyl group, a C 11 alkyl group, a C 12 alkyl group, a C 13 alkyl group, a C 14 alkyl group, a C 15 alkyl group
  • R 1 , R 2a , R 2b , R 3 , R 4a , R 4b , R 7 , R 8a , R 8b and R 9 are each independently branched or unbranched.
  • R 1 , R 2a , R 2b , R 3 , R 4a , R 4b , R 7 , R 8a , R 8b and R 9 each independently contain 0, 1, 2, 3 or 4 unsaturated bonds.
  • R 1 , R 2a , R 2b , R 3 , R 4a , R 4b , R 7 , R 8a , R 8b and R 9 are each independently substituted by an optionally substituted substituent RX .
  • R1 , R2a , R2b , R3 , R4a , R4b , R7 , R8a , R8b and R9 are each independently substituted with an optionally substituted substituent RX , and RX is selected from any of the following groups: C1 - C20 alkyl, C2 - C25 ester, C2 - C20 alkenyl, amide, urea, hydroxyl, thiol, amino.
  • the structure of the compound is as follows (as described in structure OT13-5 provided herein):
  • the compound comprises two tertiary amine groups as cation providing groups and forming a piperazine ring (p-diazepine ring) between each other, and comprises the structure (IIIa), wherein G1 and G2 are both C2 alkyl groups with a chain length of 2 atoms, and they are respectively connected to the hydrogen bond providing group urea group, and the amines of the two trivalent groups of each urea group are independently substituted, that is, R1 in the structure (IIIa) is a C12 alkyl group with a chain length of 12 atoms, R2a and R2b in the structure (IIIa) are hydrogen and C12 alkyl groups respectively (in any order), R3 in the structure (IIIa) is a C12 alkyl group, and R4a and R4b in the structure (IIIa) are hydrogen and C12 alkyl groups respectively (in any order),
  • the structure of the compound is as follows (as described in structure OT13-4 provided herein):
  • the compound comprises a tertiary amine group as a cation providing group and comprises the structure (IIIc), wherein G1 and G2 are both C6 alkyl groups with a chain length of 6 atoms, which are respectively connected to the hydrogen bond providing group urea group, and the trivalent amine directly connected to G1 or G2 of each urea group is not substituted, that is, R1 and R7 in the structure (IIIc) are hydrogen, and the trivalent amine not directly connected to G1 or G2 of each urea group is substituted independently, that is, R2a , R2b , R8a and R8b in the structure (IIIc) are all C6 alkyl groups, and R9 in the structure (IIIc) is butanol , and its molecular formula is C4H8OH .
  • the compound comprises structure (IIId):
  • G1 , G2 , G3 and G4 are is independently a linking group with a chain length of 2 to 6 atoms
  • said G 1 , G 2 , G 3 and G 4 are each independently optionally substituted
  • said H 1 and H 2 are each independently optionally substituted
  • said R 1 , R 2a , R 2b , R 7 , R 8a , R 8b , R 10 , R 11a , R 11b , R 12 , R 13a and R 13b are each independently optionally substituted.
  • G 1 , G 2 , G 3 and G 4 are each independently a linking group with a chain length of 2, 3, 4, 5, 6 atoms.
  • G 1 , G 2 , G 3 and G 4 are each independently selected from the following any group or any combination thereof: hydrocarbon group, alicyclic group, alicyclic group, alkyl group, alkenyl group, alkynyl group.
  • G1 , G2 , G3 and G4 are each independently optionally substituted, and one atom is replaced by N, replaced by O or replaced by S.
  • H1 and H2 are each independently selected from any of the following groups or any combination thereof: hydrocarbon group, alicyclic group, alicyclic heterocyclic group, alkyl group, alkenyl group, alkynyl group.
  • H1 and H2 are each independently optionally substituted, and one atom is replaced by N, replaced by O or replaced by S.
  • R 1 , R 2a , R 2b , R 7 , R 8a , R 8b , R 10 , R 11a , R 11b , R 12 , R 13a and R 13b are each independently selected from the following groups or any combination thereof: hydrocarbon group, ester group, amide group, N'-alkyl-N-alkenyl-urea group, alicyclic group, alicyclic heterocyclic group, aryl group, heteroaryl group, C 1 alkyl group, C 2 alkyl group, C 3 alkyl group, C 4 alkyl group, C 5 alkyl group, C 6 alkyl group, C 7 alkyl group, C 8 alkyl group, C 9 alkyl group, C 10 alkyl group, C 11 alkyl group, C 12 alkyl group, C 13 alkyl group, C 14 alkyl group,
  • R 1 , R 2a , R 2b , R 7 , R 8a , R 8b , R 10 , R 11a , R 11b , R 12 , R 13a and R 13b are each independently branched or unbranched.
  • R 1 , R 2a , R 2b , R 7 , R 8a , R 8b , R 10 , R 11a , R 11b , R 12 , R 13a and R 13b each independently contain 0, 1, 2, 3 or 4 unsaturated bonds.
  • R 1 , R 2a , R 2b , R 7 , R 8a , R 8b , R 10 , R 11a , R 11b , R 12 , R 13a and R 13b are each independently substituted with an optionally substituted substituent RX .
  • R 1 , R 2a , R 2b , R 7 , R 8a , R 8b , R 10 , R 11a , R 11b , R 12 , R 13a and R 13b are each independently substituted with an optionally substituted substituent RX , and RX is selected from any of the following groups: C 1 -C 20 alkyl, C 2 -C 25 ester, C 2 -C 20 alkenyl, amide, urea, hydroxyl, thiol, amino.
  • the structure of the compound is as follows (as described in structure OT13-9 provided herein): , the compound comprises two tertiary amine groups as cation providing groups, which are connected through a group with a structure of 3,6-dibutylpiperazine-2,5-dione, and the compound further comprises the structure (IIId), wherein H1 and H2 are both C4 alkyl groups, G1 , G2 , G3 and G4 are all C2 alkyl groups with a chain length of 2 atoms, and they are each connected to a hydrogen bond providing group urea group, and the trivalent amine directly connected to G1 , G2 , G3 or G4 of each urea group is not substituted, that is, R1 , R7 , R10 and R12 in the structure (IIId) are all hydrogen, and the trivalent amine not directly connected to G1 , G2 , G3 or G4 of each urea group is independently substituted, that is
  • the present application provides a compound, or a pharmaceutically acceptable salt thereof, wherein the structure of the compound is selected from any one of the structures listed in the following table:
  • any embodiment of a compound comprising structures (I), (II), or (IIIa) to (IIId) as shown above, and any specific substituents and/or variables of compound structures (I), (II), or (IIIa) to (IIId) as shown above can be independently combined with other embodiments and/or substituents and/or variables of compounds comprising structures (I), (II), or (IIIa) to (IIId) to form embodiments of the present application not specifically shown above.
  • the application provides a kind of composition
  • the composition comprises any one or more and therapeutic agent in the compound as described in the application.
  • the composition comprises any one or more, therapeutic agent and one or more excipients selected from the lipid of neutral lipid, steroid and polymer conjugation in the compound as described in the application.
  • a kind of composition of lipid nanoparticle (LNP) is provided, the LNP comprises any one, therapeutic agent and one or more excipients selected from the lipid of neutral lipid, steroid and polymer conjugation in the compound as described in the application.
  • the composition comprises any one or more, therapeutic agent and liposome in the compound as described in the application.
  • the composition also comprises other pharmaceutically acceptable excipients and/or carriers.
  • the composition comprises any one or more of the compounds described herein, a therapeutic agent, and one or more excipients selected from neutral lipids, steroids, and polymer-conjugated lipids, wherein the molar ratio of the compound to the neutral lipid is about 2: 1 to about 8: 1.
  • a composition comprising any one or more of the compounds described herein, a therapeutic agent, and one or more excipients selected from neutral lipids, steroids, and polymer-conjugated lipids
  • the compound has a molar ratio of the neutral lipid in the following numerical range: about 2: 1 to about 3: 1, about 2: 1 to about 4: 1, about 2: 1 to about 5: 1, about 2: 1 to about 6: 1, about 2: 1 to about 7: 1, about 2: 1 to about 8: 1, about 3: 1 to about 4: 1, about 3: 1 to about 5: 1, about 3: 1 to about 6: 1, about 3: 1 to about 7: 1, about 3: 1 to about 8: 1, about 4: 1 to about 5: 1, about 4: 1 to about 6: 1, About 4:1 to about 7:1, about 4:1 to about 8:1, about 5:1 to about 6:1, about 5:1 to about 7:1, about 5:1 to about 8:1, about 6:1 to about 7:1, about 6:1 to about 8:1, about 7
  • the composition comprises any one or more of the compounds as described herein, a therapeutic agent, and one or more excipients selected from neutral lipids, steroids containing cholesterol, and polymer-conjugated lipids.
  • the composition comprises any one or more of the compounds as described herein, a therapeutic agent, and one or more excipients selected from neutral lipids, steroids, and polymer-conjugated lipids, and the compound and the steroid have a molar ratio in the following numerical range: about 5:1 to about 4:1, about 5:1 to about 3:1, about 5:1 to about 2:1, about 5:1 to about 1:1, about 4:1 to about 3:1, about 4:1 to about 2:1, about 4:1 to about 1:1, about 3:1 to about 2:1, about 3:1 to about 1:1, about 2:1 to about 1:1.
  • the composition comprises any one or more of the compounds described herein, a therapeutic agent, and one or more excipients selected from a neutral lipid, a steroid, and a polymer-conjugated lipid including a pegylated lipid.
  • the composition comprises any one or more of the compounds as described herein, a therapeutic agent, and one or more excipients selected from a neutral lipid, a steroid, and a polymer-conjugated lipid, and the compound has a molar ratio to the polymer-conjugated lipid in the following numerical range: from about 100:1 to about 90:1, from about 100:1 to about 80:1, from about 100:1 to about 70:1, from about 100:1 to about 60:1, from about 100:1 to about 50:1, from about 100:1 to about 40:1, from about 100:1 to about 30:1, from about 100:1 to about 20:1, from about 90:1 to about 80:1, from about 90:1 to about 70:1, from about 90:1 to about 60:1, from about 90:1 to about 50:1, from about 90:1 to about 40:1, from about 90:1 to about 30:1, about 90:1 to about 20:1, about 80:1 to about 70:1, about 80:1 to about 70:1, about 80:1 to
  • the diameter of the composition is about 500nm or less.
  • the composition has a diameter of the following values: about 500nm, about 450nm, about 400nm, about 350nm, about 300nm, about 250nm, about 200nm, about 150nm, about 100nm, about 75nm, about 50nm, about 25nm.
  • the therapeutic agent comprises one or more nucleic acids.
  • the content disclosed in this application is not limited to the specific nucleic acids disclosed in this specification, and is not limited in scope to any specific source, sequence or type of nucleic acid, but a person of ordinary skill in the art can easily identify related homologs in various other nucleic acid sources, including nucleic acids from non-human species. It is contemplated that the nucleic acids used in the content disclosed in this application may comprise sequences based on naturally occurring sequences.
  • the therapeutic agent comprises one or more nucleic acids
  • the nucleic acid comprises one or more modified nucleosides (for example, comprising one or more nucleosides modified with a sugar moiety), in the hope that the compound comprising the nucleic acid may have desired properties.
  • the therapeutic agent comprises one or more nucleic acids that are oligonucleotides comprising modified nucleosides (e.g., oligonucleotides comprising modified sugars, modified nucleobases, modified bonds, and/or modified nucleotides at other positions such as the 3' position of the sugar on the 3' terminal nucleotide and/or the 5' position of the 5' terminal nucleotide), and the oligonucleotides obtained by selecting a particular modification may have desired characteristics.
  • modified nucleosides e.g., oligonucleotides comprising modified sugars, modified nucleobases, modified bonds, and/or modified nucleotides at other positions such as the 3' position of the sugar on the 3' terminal nucleotide and/or the 5' position of the 5' terminal nucleotide
  • the therapeutic agent comprises components of the CRISPR system and/or nucleic acids encoding Cas enzymes.
  • the "CRISPR system” may also be referred to as a “CRISPR/Cas system", which is a tool for site-specific genome targeting in an organism.
  • CRISPR/Cas system may be a type II CRISPR/Cas system, which is a prokaryotic adaptive immune response system that uses non-coding RNA to guide Cas nucleases to induce site-specific DNA cleavage, and subsequently triggers cellular DNA repair mechanisms, i.e., repairing DNA damage through non-homologous end-joining DNA repair pathways (NHEJ) or homology-directed repair (HDR) pathways; the system may also be delivered to mammalian cells or eukaryotic cells to mediate genome editing (e.g., gene knockout or knock-in).
  • NHEJ non-homologous end-joining DNA repair pathways
  • HDR homology-directed repair
  • the therapeutic agent can include one or more Cas nucleases, non-limiting examples of which include Cas1, Cas1B, Cas2, Cas3, Cas4, Cas5, Cas6, Cas7, Cas8, Cas9 (also known as Csn1 and Csx12), Cas10, Csy1, Csy2, Csy3, Cse1, Cse2, Csc1, Csc2, Csa5, Csn2, Csm2, Csm3, Csm4 , Csm5, Csm6, Cmr1, Cmr3, Cmr4, Cmr5, Cmr6, Csb1, Csb2, Csb3, Csx17, Csx14, Csx10, Csx16, CsaX, Csx3, Csx1, Csx15, Csf1, Csf2, Csf3, Csf4, homologs thereof or modified forms thereof, these enzymes are known and can be easily found by a person skilled in the art from, for example, the SwissPro
  • the therapeutic agent may comprise a Cas9 enzyme and/or a mutant form thereof for specific binding and/or site-directed cleavage of a single-stranded or double-stranded target polynucleotide at a target sequence position.
  • the therapeutic agent comprises a nucleic acid encoding a component of a CRISPR system, wherein “encoding” means that the target polynucleotide sequence can be transcribed and/or translated according to a genetic code that functions in a host cell/host animal to produce a corresponding gene product (e.g., a nuclease).
  • the therapeutic agent comprises a guide RNA (guide RNA or gRNA).
  • the therapeutic agent comprises a nucleic acid encoding a Cas enzyme.
  • the "nucleic acid encoding a Cas enzyme” comprises DNA (e.g., a plasmid) or RNA (e.g., mRNA).
  • the present application provides a cell comprising a compound and/or composition as described in the present application.
  • the cell is a carrier cell comprising a negatively charged cell membrane.
  • the LNP comprising the compound described in the present application is protonated under a suitable pH environment, and then aggregated with the cell membrane of the carrier cell through electrostatic interaction.
  • the compound and/or composition described in the present application is protonated under a suitable pH environment, so that the cell membrane of the carrier cell mediates endocytosis or membrane permeation to include the compound and/or composition.
  • the present application provides a kit, the kit comprising the compound described in the present application, The composition and/or the cell described in the present application.
  • the kit includes at least one vial, tube, flask, bottle, syringe or other suitable container, in which the components can be placed.
  • the kit will also include a second, third or other additional container, in which the additional components can be placed separately.
  • various combinations of components can be included in the container.
  • all LNP components are combined in a single container.
  • some or all of the LNP components are provided in separate containers.
  • the kit will also include packaging for accommodating various containers.
  • the kit includes instructions for using the kit components.
  • the instructions include a paper-based physical form and/or a machine-readable electronic form.
  • the present application provides a method of administering a therapeutic agent, the method comprising providing a compound described herein, a composition described herein, a cell described herein, and/or a kit described herein.
  • the method comprises mixing the therapeutic agent described herein with the compound described herein.
  • the method comprises placing the therapeutic agent described herein in a composition described herein, a cell described herein, and/or a kit described herein.
  • the present application provides a method for delivering nucleic acid, the method comprising providing a compound described in the present application, a composition described in the present application, a cell described in the present application, and/or a kit described in the present application.
  • the method comprises mixing the nucleic acid described in the present application with the compound described in the present application.
  • the method comprises placing the nucleic acid described in the present application in a composition described in the present application, a cell described in the present application, and/or a kit described in the present application.
  • the method for delivering nucleic acid comprises delivering the compound described in the present application, the composition described in the present application, the cell described in the present application, and/or the kit described in the present application to the whole body, so that it is widely exposed to most of the body, and can be carried out by any means known in the art, including but not limited to intravenous, intra-arterial, subcutaneous and intraperitoneal delivery.
  • the method for delivering nucleic acid comprises delivering the compound described in the present application, the composition described in the present application, the cell described in the present application, and/or the kit described in the present application to the local area, so that it directly reaches the target site in the organism, and can be carried out by, for example, direct injection into the disease site (such as, tumor or inflammation site) or target organ (such as, liver, heart, pancreas, kidney, etc.).
  • the local delivery includes local administration or local injection technology, including but not limited to intramuscular, subcutaneous or intradermal injection. For example, such local delivery does not preclude systemic pharmacological effects.
  • the present application provides a pharmaceutical composition, which comprises the compound described in the present application, the composition described in the present application, the cell described in the present application and/or the kit described in the present application.
  • the pharmaceutical composition further comprises a pharmaceutically acceptable auxiliary component.
  • the pharmaceutical composition is used to treat and/or prevent a disease. sick.
  • the present application provides the use of the compounds described herein, the compositions described herein, the cells described herein, the kits described herein and/or the pharmaceutical compositions described herein for preparing drugs for treating or preventing diseases.
  • the present application provides the compounds described herein, the compositions described herein, the cells described herein, the kits described herein and/or the pharmaceutical compositions described herein, which are used for treating or preventing diseases.
  • the present application provides a method for treating and/or preventing a disease, which comprises administering to a subject or patient a suitable dose of the compound described herein, the composition described herein, the cell described herein and/or the kit described herein.
  • the disease comprises cancer.
  • the diseases include diseases associated with protein or enzyme deficiency.
  • the disease includes diseases associated with a specific miRNA or a group or more than one group of miRNAs.
  • Aqueous hydrazine was added to ethanol and reacted with 2-[(10E,12E)-octadeca-9,12-dienyl]isoindole-1,3-dione (the product of step 6) under inert gas at 0-20° C. for two hours.
  • the product was dried and purified by silica gel column chromatography.
  • 19-Ethoxy-9,11-bis(ethoxycarbonyl)-10,19-dioxyylidene nonadecanoic acid ethyl ester (the product of step 10.) was reacted with 12M concentrated hydrochloric acid and acetic acid at 110°C for more than 12 hours to obtain a product which was washed with a saturated sodium chloride solution, dried over anhydrous sodium sulfate, and the remaining solvent was removed by distillation under reduced pressure to obtain a product which was then purified by silica gel chromatography.
  • 10-Oxylidene nonadecanedioic acid product of step 11.
  • 2-butyloctanol were subjected to esterification reaction at 30°C for 12 hours under the catalysis of EDCI, DMAP and DCM.
  • the organic layer was separated and washed with saturated sodium chloride solution, and the remaining solvent was removed under reduced pressure to obtain the product, which was purified by silica gel chromatography.
  • 6-Bromohexan-1-amine (the product of step 15) was heated to 70°C in THF with dihexylamine under the catalysis of CDI, and neopentane was introduced and the reaction was shaken for 2 hours.
  • the product was purified by silica gel column chromatography.
  • CDI N,N'-carbonyldiimidazole
  • 2-[4-(2-aminoethyl)piperazin-1-yl]ethan-1-amine was catalyzed by CDI and heated to 70°C in THF, and then introduced with neopentane and shaken for 2 hours.
  • the product was purified by silica gel column chromatography to obtain a light yellow solid.
  • OT13-6 is the same as the synthesis method described in step 19.
  • 1-(Dodecylamino)-N- ⁇ 2-[4-(4-oxyylidene-3,5-diazaheptadecane-1-yl)piperazine-1-yl]ethyl ⁇ methaneamide (the product of step 18.) and 12-oxirane are mixed, sealed in THF containing chloroacetonitrile, heated at 80°C and stirred for 2 days until the target product is detected by MALDI-TOF mass spectrometry.
  • the crude product is purified by silica gel chromatography, and the product is finally confirmed by nuclear magnetic resonance.
  • OT13-7 is the same as the synthesis method described in step 19.
  • the reactant is replaced with an unsaturated chain 2-[(6E)-dode-6-enyl] oxirane in step 18.
  • the mixture is sealed in THF containing chloroacetonitrile and heated at 80°C for 2 hours with continuous stirring.
  • the reaction was continued for 14 days until the target product was detected by MALDI-TOF mass spectrometry.
  • the crude product was purified by silica gel chromatography and the product was finally confirmed by NMR.
  • 1,4-bis(2-hydroxyethyl)piperazine is dissolved in a mixed system of DMF and acetonitrile, 2-bromoethylamine hydrobromide is added, and the mixture is reacted at 50-100°C for six hours under acidic conditions and catalyzed by potassium bromide.
  • Benzyl ⁇ [9-amino-12-(methoxycarbonyl)-3,10-dioxyylidene-1-phenyl-4,11-diaza-2-oxahexadecan-16-yl]amino ⁇ methane (the product of step 26) was dissolved in methanol and reacted at 25° C. for 12 hours in an alkaline environment (adjusted by sodium hydroxide). The product was washed with water and dried in vacuo.
  • step 32 The product of step 32. is subjected to hydrogenation reaction in an ethanol reaction system under the catalysis of Lindela catalyst to generate 2- ⁇ [(4Z)-undec-4-enyl]oxy ⁇ tetrahydropyran, and the product is purified by silica gel column chromatography.
  • p-Toluenesulfonic acid is dissolved in methanol and reacted with the product of step 33 to generate (4Z)-undec-4-en-1-ol.
  • the product is purified by silica gel column chromatography.
  • step 35 The product of step 35. is reacted with sodium azide in the presence of DMSO, and the resulting product is purified by silica gel column chromatography.
  • step 36 The product of step 36. is subjected to hydrogenation substitution reduction in THF solvent by catalysis of triphenylphosphine hydrate.
  • the product is purified by silica gel chromatography and used as
  • [(4-Bromobutyl)amino]methane acid-2-methylpropan-2-yl ester is reacted with 2-oxo-pyridinium-1-carboxylic acid-2-methylpropan-2-yl ester in a THF reaction solvent containing magnesium powder and iodine, and the product is purified by silica gel column chromatography.
  • Sodium cyanoborohydride was added to a mixed reaction system of ethanol and ammonium acetate at room temperature and stirred for more than 2 hours. The product was filtered through silica and washed with hot ethyl acetate.
  • step 40 The product of step 40. was deprotected from Boc in a mixed system of ethyl acetate/hydrochloric acid, and the product was purified by silica gel chromatography to obtain the product, which was then heated to 70° C. with (4Z)-undec-4-en-1-amine (product of step 37.) in THF, and neopentane was introduced and shaken for 2 hours. The product was purified by silica gel chromatography to obtain OT13-10.
  • step 42 The product of step 42. was reacted with 2-methylpropan-2-yl [(5-formylpentyl)amino]methanoate in methanol at room temperature by adding sodium cyanoborohydride. The stirring was continued for more than 2 hours. The product was filtered through silica and washed with hot ethyl acetate.
  • step 43 is deprotected from Boc in a mixed system of ethyl acetate/hydrochloric acid; in addition, heptyl-1-amine and 1-iodohexane are reacted in an acetonitrile solvent containing potassium carbonate to generate the product hexyl (octyl) amine, which is purified by a silica gel column and used in the next reaction.
  • the product obtained after de-Bocation of the product in step 43. is heated to 70° C. in THF with hexyl(octyl)amine under the catalysis of CDI, and neopentane is introduced to react with shaking for 2 hours.
  • the product is purified by silica gel column chromatography to obtain the product.
  • [(5-Bromopentyl)amino]methane acid-2-methylprop-2-yl ester reacts with 2-(benzyloxy)ethyl-1-amine in an acetonitrile solvent containing potassium carbonate at room temperature to generate a product, which is purified by silica gel column chromatography.
  • step 47 benzyl [(3-formylpropyl)amino]methanoate was added at room temperature, and the mixture was stirred for more than 2 hours under the catalysis of sodium cyanoborohydride. The product was filtered through silica and washed with hot ethyl acetate for later use.
  • step 48 The product of step 48 was de-Boc protected in an ethyl acetate/hydrochloric acid mixture system, and the product was purified by silica gel column chromatography and used in the next reaction.
  • 1-Bromooctane is reacted with decanoic acid in THF solvent under the catalysis of sodium hydride at room temperature under continuous stirring for two hours to generate 2-octyldecanoic acid, and the product is purified by silica gel chromatography column for use.
  • 2-Octyldecanoic acid was dissolved in THF solvent and stirred continuously at room temperature for 2 h.
  • 2-Octyldecan-1-ol was generated by LaN-catalyzed reduction and the product was purified by silica gel column chromatography.
  • step 52 is subjected to Gabriel primary amine reaction, the reaction system is ethanol, and it reacts with hydrazine hydrate at 80° C. to generate 2-octyldec-1-amine, and the product is purified by silica gel column chromatography.
  • step 53 The product 2-octyldec-1-amine obtained in step 53 and the product in step 49 are heated to 70° C. in THF, and new pentane is introduced. The mixture is shaken for 2 hours to obtain the product, which is then purified via silica gel chromatography.
  • a dry round-bottom flask is filled with hydrogen, and the product of step 54 is added to a dichloromethane solvent containing acetic acid, and reacted at room temperature for 2 hours under the catalysis of Pd or C to perform debenzenization reduction.
  • the reaction needs to be maintained at a pressure of 500 psi and is achieved in a stainless steel pressure vessel.
  • the product is filtered, concentrated by rotary evaporation, and purified by silica gel chromatography.
  • Step 55 The obtained product was reacted with undecane-1-amine in THF at 70°C, and neopentane was introduced, and the reaction was shaken for 2 hours to obtain OT13-12.
  • the product was purified by silica gel chromatography.
  • LNP nanoparticles are prepared to achieve effective encapsulation of gene editing tools or other nucleic acid drugs.
  • the organic phase contains at least one ionizable lipid (OT13-1 to OT13-9), at least one supporting lipid, at least one amphiphilic block copolymer and cholesterol, and is dissolved in an organic solvent that is miscible with water.
  • the organic solvent is preferably selected from ethanol, acetonitrile, acetone, etc.
  • OT13-3, DSPC, PEG-DMG, and cholesterol are used, wherein the four components The molar ratio is 50:10:1.5:38.5;
  • Aqueous phase an aqueous solution of a gene editing tool, wherein the nucleic acid material uses yeast extracted RNA as a template RNA (McLean, log. R822593), the content is 0.5-50% (w/v) (the preferred content in this embodiment is 40% (w/v)), and the pH is about 3.0 to about 7.0 (the preferred pH in this embodiment is about 4.0).
  • the aqueous salt solution can be selected from: citric acid buffer, phosphate buffer, Tris-HCl buffer system; wherein, the citric acid buffer is used in this embodiment.
  • the mixing of organic and aqueous phases can be achieved by microfluidics and impinging flow reactors.
  • the embedding efficiency of gene editing tool RNA can be optimized by regulating the N/P ratio of the system, which ranges from 1:1 to 9:1.
  • LNP was prepared by the method of this example, and the particle size, dispersion (PDI), and embedding efficiency (EE) were tested. The results are shown in the following table:
  • the embedding rate characterization method the embedding rate of the prepared LNP is measured by Quant-iTTM RNA Reagent and Kit. Specific detection method:
  • Solution preparation Dilute an appropriate amount of 20 ⁇ TE buffer to 1 ⁇ with ultrapure water, enough for the experiment on the day. Dilute the concentrated dye solution with 1 ⁇ TE (large range 25-1000ng/ml RNA) at a ratio of 1:200, (small range 1-50ng/ml RNA) at a ratio of 1:2000. Wrap it with foil or store it in a dark place away from light. Dilute TritonX-100 with 1 ⁇ TE buffer to a concentration of 5% for later use.
  • Sample processing Set ultrapure water with RNA concentration of 0 as blank background, set LNP with TE buffer instead of 5% TritonX-100 as free RNA determination sample, set LNP with 5% TritonX-100 as total RNA determination sample. Take the LNP sample to be determined and the blank and add equal volumes of 5% TritonX-100 and 1 ⁇ TE solution, incubate at 50-60°C for 5-10 minutes. The incubated LNP sample is diluted to 10-400 times with 1 ⁇ TE buffer (appropriately adjusted according to the RNA concentration of the sample group). After dilution, take 100 ⁇ l and add it to a 96-well plate, and then add 100 ⁇ l of the diluted concentrated dye solution in the dark, and measure the fluorescence absorbance within 5 minutes. The determination conditions are excitation wavelength 480nm, emission wavelength 520nm.
  • LNP was prepared by the method of Example 2, wherein the ionizable lipid was OT13-3 and the embedded nucleic acid was Luciferase mRNA (nucleotide sequence is shown in the following table).
  • the prepared Luc-mRNA-LNP was injected into 6-8 week old C57BL/6 mice, and the expression level of Lurciferase in the mouse liver was observed by in vivo imaging technology 6 hours after injection ( Figure 1), so as to evaluate Delivery effect of OT13 lipid-formed LNP in mouse liver.
  • Figure 1 so as to evaluate Delivery effect of OT13 lipid-formed LNP in mouse liver.
  • the results showed that 6 hours after injection, LNP containing ionizable lipid OT13-3 successfully delivered Luc-mRNA to mouse liver and achieved efficient Luciferase expression.
  • the LNP containing OT13-3 ionizable lipids was prepared in the manner of Example 2, and a sample of Cre mRNA (nucleotide sequence is shown in the table below) was embedded at the same time. It was injected into adult Ai9 transgenic mice. Five days after injection, the mouse liver was removed for frozen sectioning and observed with a laser confocal microscope. The results showed that LNP containing OT13-3 can effectively transfect and edit the adult mouse liver ( Figure 2).
  • PCSK9 is a target gene suitable for analyzing the delivery of functional gene editing tools to the liver. It is produced in liver cells and secreted into the blood circulation, so it can be used for in vivo screening of liver-directed lipid molecules.
  • EPIREG-mRNA and gRNA the complementary nucleotide sequence of its targeting sequence is shown in the following table
  • a gene editing tool with a silencing effect for PCSK9 are co-encapsulated into LNPs containing OT13-3 by the method of Example 2, and injected into wild-type 6-8-week C57BL/6 mice at a concentration of 3 mg/kg RNA. After 9 days, the kit was used for PCSK9 detection (Figure 3).
  • OT13-3 forms LNPs with nearly 80% epigenetic editing and silencing of target genes.
  • the epigenetic editing efficacy of OT13-3-formed LNPs in vivo can be compared with a positive control, which is commercially applied to LNPs formed by ionizable lipids for clinical use.
  • the evaluation data of this embodiment proves that the OT13 series of molecules can be used as a nucleic acid delivery system in the field of liver-targeted gene therapy.

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Abstract

Provided in the present application are a compound for a delivery system and the use thereof, and an ionizable lipid having a hydrogen bond providing group.

Description

一种用于递送系统的化合物及其用途A compound for delivery system and its use 技术领域Technical Field
本申请涉及生物医药领域，具体的涉及一种用于递送系统的化合物及其用途。The present application relates to the field of biomedicine, and specifically to a compound for a delivery system and its use.
背景技术Background technique
基于可翻译核苷酸递送的治疗具有巨大的临床应用潜力，其治疗过程是通过合成构建体以产生可供选择的核酸表达产物——蛋白质序列，且无论该序列是否是系统固有的，都可以达到增加蛋白质的现有水平、替代蛋白质的缺失或无功能形式、以及在细胞或有机体中引入新的蛋白质及其相关生物功能的效果。治疗性核酸包括如小型干扰RNA(siRNA)、信使RNA(mRNA)、反义寡核苷酸、核酶、DNA酶、质粒、免疫刺激性核酸、antagomir、antimiR、mRNA模拟物、supermir.UI衔接子和适配体，使它们在特定细胞中表达在治疗例如与蛋白质或酶缺乏有关的疾病中是至关重要的，因此将这些治疗性核酸有效地递送至细胞或有机体内的目标位点是实现基于核酸的治疗的关键挑战。Therapies based on the delivery of translatable nucleotides have great potential for clinical applications. The therapeutic process is to produce selectable nucleic acid expression products - protein sequences through synthetic constructs, and whether the sequence is inherent in the system, it can achieve the effect of increasing the existing level of protein, replacing the missing or non-functional form of protein, and introducing new proteins and their related biological functions in cells or organisms. Therapeutic nucleic acids include small interfering RNA (siRNA), messenger RNA (mRNA), antisense oligonucleotides, ribozymes, DNA enzymes, plasmids, immunostimulatory nucleic acids, antagomirs, antimiRs, mRNA mimics, supermir.UI adapters and aptamers. Their expression in specific cells is crucial in the treatment of diseases such as those related to protein or enzyme deficiency. Therefore, the effective delivery of these therapeutic nucleic acids to target sites in cells or organisms is a key challenge to achieve nucleic acid-based therapy.
在当前的治疗环境中使用寡核苷酸构建体主要面临着两方面的问题。首先，游离RNA本身结构的不稳定性使得其在递送途中极易被体内的多种酶迅速降解；其次，游离RNA进入相关翻译系统所在的细胞内区室的能力有限。该限制具体表现为：其一，带有负电荷的RNA骨架与细胞膜表面的负电荷产生静电排斥所导致的膜屏障；其二，构建体通过与细胞膜静电结合产生的内吞作用进入细胞后将被包裹在胞内体(endosome)中，成熟的胞内体随后将与包含有特定消化酶的溶酶体融合，因而构建体需要从胞内体和/或溶酶体中逃逸出来以防止引发构建体的负载核酸被降解。由脂质与其他脂质组分(如中性脂质、类固醇、聚乙二醇化脂质)共同配制的、用于装载寡核苷酸的脂质纳米颗粒(LNP)已经用于阻断体内RNA的降解并促进细胞对递送的寡核苷酸的摄取，但仍然需要研究用于递送寡核苷酸的优化脂质和LNP，以提高其封装效率、内容药物的完整性、逃逸率和LNP相关药物配方的安全性等性能。The use of oligonucleotide constructs in the current therapeutic environment mainly faces two problems. First, the instability of the structure of free RNA itself makes it extremely easy to be rapidly degraded by various enzymes in the body during delivery; second, the ability of free RNA to enter the intracellular compartment where the relevant translation system is located is limited. This limitation is specifically manifested in the following ways: first, the negatively charged RNA skeleton and the negative charge on the cell membrane surface produce electrostatic repulsion, resulting in a membrane barrier; second, after the construct enters the cell through endocytosis generated by electrostatic binding to the cell membrane, it will be encapsulated in the endosome, and the mature endosome will then fuse with the lysosome containing specific digestive enzymes, so the construct needs to escape from the endosome and/or lysosome to prevent the degradation of the load nucleic acid that triggers the construct. Lipid nanoparticles (LNPs) co-formulated with lipids and other lipid components (e.g., neutral lipids, steroids, PEGylated lipids) for loading oligonucleotides have been used to block RNA degradation in vivo and promote cellular uptake of delivered oligonucleotides, but there is still a need to study optimized lipids and LNPs for delivering oligonucleotides to improve their encapsulation efficiency, integrity of the content drug, escape rate, and safety of LNP-related drug formulations.
发明内容Summary of the invention
本申请提供了一种用于递送系统的化合物，包括其立体异构体、药学上可接受的盐、前药或互变异构体；以及其用途，包括单独使用该化合物，或者与其他脂质组分如中性脂质、带电脂质、类固醇和/或其类似物、和/或聚合物缀合的脂质组合使用以形成脂质纳米颗粒以用 于递送治疗剂。本申请的化合物在可响应pH值变化而发生质子化的头部基团(例如叔胺)附近2-6个原子的位置处设计了可与RNA碱基形成分子间氢键的取代基团(例如脲基)。该设计形成的与核酸间的弱相互作用力与静电作用协同装载RNA，可以提升LNP或脂质体(liposome)的封装效率(Encapsulation efficiency)，同时提升脂质和装载药物的完整性(Lipid and cargo integrity)。此外，带有脲基的可电离脂质与现有技术相比(例如ALCO315),由于羟基的亲电子性被两边的胺中和掉而不会与叔胺形成分子内六元环或五元环，更有利于加快LNP在体内降解，从而提升包含有该LNP的配方安全性。The present application provides a compound for a delivery system, including a stereoisomer, a pharmaceutically acceptable salt, a prodrug or a tautomer thereof; and its use, including using the compound alone, or using it in combination with other lipid components such as neutral lipids, charged lipids, steroids and/or their analogs, and/or polymer-conjugated lipids to form lipid nanoparticles for use For delivering therapeutic agents. The compounds of the present application are designed with a substituent group (e.g., urea group) that can form an intermolecular hydrogen bond with the RNA base at the position of 2-6 atoms near the head group (e.g., tertiary amine) that can be protonated in response to changes in pH. The weak interaction force between the nucleic acid formed by this design and the electrostatic effect synergistically load RNA, which can improve the encapsulation efficiency (Encapsulation efficiency) of LNP or liposomes (liposomes), while improving the integrity (Lipid and cargo integrity) of lipids and loaded drugs. In addition, compared with the prior art (e.g., ALCO315), the ionizable lipids with urea groups are more conducive to accelerating the degradation of LNP in vivo, thereby improving the safety of the formulation containing the LNP, because the electrophilicity of the hydroxyl group is neutralized by the amines on both sides and does not form an intramolecular six-membered ring or five-membered ring with the tertiary amine.
一方面，本申请提供了一种化合物，或其药学上可接受的盐，其包含阳离子提供基团以及氢键提供基团，且所述阳离子提供基团与氢键提供基团之间包含2至6个原子。In one aspect, the present application provides a compound, or a pharmaceutically acceptable salt thereof, comprising a cation providing group and a hydrogen bond providing group, and the cation providing group and the hydrogen bond providing group contain 2 to 6 atoms.
在一些实施方案中，所述化合物包含阳离子脂质。In some embodiments, the compound comprises a cationic lipid.
在一些实施方案中，所述化合物包含可电离脂质。In some embodiments, the compound comprises an ionizable lipid.
在一些实施方案中，所述阳离子提供基团包含在pH为7.0约或更低的条件下带正电的基团。In some embodiments, the cation providing group comprises a group that is positively charged at a pH of about 7.0 or less.
在一些实施方案中，所述阳离子提供基团包含在pH为4.0约或更低的条件下带正电的基团。In some embodiments, the cation providing group comprises a group that is positively charged at a pH of about 4.0 or less.
在一些实施方案中，所述阳离子提供基团包含叔胺基团。In some embodiments, the cation-providing group comprises a tertiary amine group.
在一些实施方案中，所述氢键提供基团包含脲基。In some embodiments, the hydrogen bond providing group comprises a urea group.
在一些实施方案中，所述化合物包含尾部，其具有疏水性。In some embodiments, the compound comprises a tail that is hydrophobic.
在一些实施方案中，所述尾部包含饱和和/或不饱和的脂肪链。In some embodiments, the tail comprises a saturated and/or unsaturated fatty chain.
在一些实施方案中，所述尾部取代在所述脲基的一个或两个氮原子上。In some embodiments, the tail is substituted on one or both nitrogen atoms of the urea group.
在一些实施方案中，所述尾部的主链链长为1至20个原子。In some embodiments, the backbone chain length of the tail is 1 to 20 atoms.
在一些实施方案中，所述化合物包含以下结构(I)：
In some embodiments, the compound comprises the following structure (I):
其中，L1为氢键提供基团，G1为链长为2至6个原子的连接基团，所述L1任选地被取代，所述G1任选地被取代。Wherein, L1 is a hydrogen bond providing group, G1 is a linking group with a chain length of 2 to 6 atoms, said L1 is optionally substituted, and said G1 is optionally substituted.
在一些实施方案中，所述化合物包含以下结构(II)：
In some embodiments, the compound comprises the following structure (II):
其中，G1为链长为2至6个原子的连接基团，所述G1任选地被取代，R1、R2a和R2b各自独立地选自任选取代的取代基。Wherein, G1 is a linking group having a chain length of 2 to 6 atoms, said G1 is optionally substituted, and R1 , R2a and R2b are each independently selected from an optionally substituted substituent.
在一些实施方案中，所述化合物包含以下结构(IIIa)：
In some embodiments, the compound comprises the following structure (IIIa):
其中，G1和G2各自独立地为链长为2至6个原子的连接基团，所述G1和G2各自独立地任选地被取代，R1、R2a、R2b、R3、R4a和R4b各自独立地选自任选取代的取代基。wherein G1 and G2 are each independently a linking group having a chain length of 2 to 6 atoms, said G1 and G2 are each independently optionally substituted, and R1 , R2a , R2b , R3 , R4a and R4b are each independently selected from optionally substituted substituents.
在一些实施方案中，所述化合物包含以下结构(IIIb)：
In some embodiments, the compound comprises the following structure (IIIb):
其中，所述G1为链长为2至6个原子的连接基团，所述G1任选地被取代，R1、R2a、R2b、R5和R6各自独立地选自任选取代的取代基。Wherein, the G1 is a linking group with a chain length of 2 to 6 atoms, the G1 is optionally substituted, and R1 , R2a , R2b , R5 and R6 are each independently selected from optionally substituted substituents.
在一些实施方案中，所述化合物包含以下结构(IIIc)：
In some embodiments, the compound comprises the following structure (IIIc):
其中，G1和G2各自独立地为链长为2至6个原子的连接基团，所述G1和G2各自独立地任选地被取代，R1、R2a、R2b、R7、R8a、R8b和R9各自独立地选自任选取代的取代基。wherein G1 and G2 are each independently a linking group having a chain length of 2 to 6 atoms, said G1 and G2 are each independently optionally substituted, and R1 , R2a , R2b , R7 , R8a , R8b and R9 are each independently selected from optionally substituted substituents.
在一些实施方案中，所述化合物包含以下结构(IIId)：
In some embodiments, the compound comprises the following structure (IIId):
其中，G1、G2、G3和G4各自独立地为链长为2至6个原子的连接基团，所述G1、G2、G3和G4各自独立地任选地被取代，所述H1和H2各自独立地任选地被取代，R1、R2a、R2b、R7、R8a、R8b、R10、R11a、R11b、R12、R13a和R13b各自独立地选自任选取代的取代基。wherein G1 , G2 , G3 and G4 are each independently a linking group having a chain length of 2 to 6 atoms, said G1 , G2 , G3 and G4 are each independently optionally substituted, said H1 and H2 are each independently optionally substituted, and R1 , R2a , R2b , R7 , R8a , R8b , R10 , R11a , R11b , R12 , R13a and R13b are each independently selected from optionally substituted substituents.
在一些实施方案中，所述G1、G2、G3和G4各自独立地为烃基、脂环基、和/或脂杂环基。In some embodiments, G 1 , G 2 , G 3 and G 4 are each independently a hydrocarbon group, an alicyclic group, and/or an alicyclic heterocyclic group.
在一些实施方案中，所述G1、G2、G3和G4各自独立地为烷基、烯基、和/或炔基。In some embodiments, G 1 , G 2 , G 3 and G 4 are each independently alkyl, alkenyl, and/or alkynyl.
在一些实施方案中，所述G1、G2、G3和G4中的一个原子各自独立地被N、O或S替换。In some embodiments, one atom in G 1 , G 2 , G 3 and G 4 is each independently replaced by N, O or S.
在一些实施方案中，所述H1和H2各自独立地为烃基、脂环基、和/或脂杂环基。In some embodiments, H 1 and H 2 are each independently a hydrocarbon group, an alicyclic group, and/or an alicyclic heterocyclic group.
在一些实施方案中，所述H1和H2各自独立地为烷基、烯基、和/或炔基。In some embodiments, H 1 and H 2 are each independently alkyl, alkenyl, and/or alkynyl.
在一些实施方案中，所述H1和H2中的一个原子各自独立地被N、O或S替换。In some embodiments, one atom in H1 and H2 is each independently replaced by N, O or S.
在一些实施方案中，所述R1、R2a、R2b、R3、R4a、R4b、R5、R6、R7、R8a、R8b、R9、R10、R11a、R11b、R12、R13a、和R13b各自独立地为烃基、酯基、酰胺、N’-烷基-N-烯基-脲基、脂环基、脂杂环基、芳基、和/或杂芳基。In some embodiments, R 1 , R 2a , R 2b , R 3 , R 4a , R 4b , R 5 , R 6 , R 7 , R 8a , R 8b , R 9 , R 10 , R 11a , R 11b , R 12 , R 13a , and R 13b are each independently a hydrocarbon group, an ester group, an amide group, an N′-alkyl-N-alkenyl-urea group, an alicyclic group, an alicyclic heterocyclic group, an aryl group, and/or a heteroaryl group.
在一些实施方案中，所述R1、R2a、R2b、R3、R4a、R4b、R5、R6、R7、R8a、R8b、R9、R10、R11a、R11b、R12、R13a、和R13b各自独立地为C1-C20烷基、C2-C40烯基、C2-C45酯基和/或C2-C20炔基。In some embodiments, R 1 , R 2a , R 2b , R 3 , R 4a , R 4b , R 5 , R 6 , R 7 , R 8a , R 8b , R 9 , R 10 , R 11a , R 11b , R 12 , R 13a , and R 13b are each independently C 1 -C 20 alkyl, C 2 -C 40 alkenyl, C 2 -C 45 ester and/or C 2 -C 20 alkynyl.
在一些实施方案中，所述R1、R2a、R2b、R3、R4a、R4b、R5、R6、R7、R8a、R8b、R9、R10、R11a、R11b、R12、R13a、和R13b各自独立地为支化的或非支化的。In some embodiments, R 1 , R 2a , R 2b , R 3 , R 4a , R 4b , R 5 , R 6 , R 7 , R 8a , R 8b , R 9 , R 10 , R 11a , R 11b , R 12 , R 13a , and R 13b are each independently branched or unbranched.
在一些实施方案中，所述R1、R2a、R2b、R3、R4a、R4b、R5、R6、R7、R8a、R8b、R9、R10、 R11a、R11b、R12、R13a、和R13b各自独立地包含0至4个不饱和键。In some embodiments, R 1 , R 2a , R 2b , R 3 , R 4a , R 4b , R 5 , R 6 , R 7 , R 8a , R 8b , R 9 , R 10 , R 11a , R 11b , R 12 , R 13a , and R 13b each independently contain 0 to 4 unsaturated bonds.
在一些实施方案中，所述R1、R2a、R2b、R3、R4a、R4b、R5、R6、R7、R8a、R8b、R9、R10、R11a、R11b、R12、R13a、和R13b各自独立地被Rx取代，所述Rx为任选取代的取代基。在某些实施方案中，所述RX为C1-C20烷基、C2-C25酯基、C2-C20烯基、酰胺、脲基、羟基、巯基或氨基。In some embodiments, R 1 , R 2a , R 2b , R 3 , R 4a , R 4b , R 5 , R 6 , R 7 , R 8a , R 8b , R 9 , R 10 , R 11a , R 11b , R 12 , R 13a , and R 13b are each independently substituted by R x , and R x is an optionally substituted substituent. In certain embodiments, R x is C 1 -C 20 alkyl, C 2 -C 25 ester, C 2 -C 20 alkenyl, amide, urea, hydroxyl, sulfhydryl or amino.
另一方面，本申请提供了一种化合物，或其药学上可接受的盐，所述化合物选自如本申请所提供的结构。On the other hand, the present application provides a compound, or a pharmaceutically acceptable salt thereof, wherein the compound is selected from the structures provided in the present application.
另一方面，本申请提供了一种组合物，其包含所述的化合物以及治疗剂。On the other hand, the present application provides a composition comprising the compound and a therapeutic agent.
在一些实施方案中，所述组合物还包含选自以下组的物质：中性脂质、类固醇以及聚合物缀合的脂质。In some embodiments, the composition further comprises a substance selected from the group consisting of a neutral lipid, a steroid, and a polymer-conjugated lipid.
在一些实施方案中，所述组合物包含脂质体。In some embodiments, the composition comprises liposomes.
在一些实施方案中，所述组合物直径约为500nm或更小。In some embodiments, the composition is about 500 nm or less in diameter.
在一些实施方案中，所述类固醇包含胆固醇。In some embodiments, the steroid comprises cholesterol.
在一些实施方案中，所述聚合物缀合的脂质包含聚乙二醇化脂质。In some embodiments, the polymer-conjugated lipid comprises a PEGylated lipid.
在一些实施方案中，所述化合物与所述中性脂质的摩尔比为约2:1至约8:1.In some embodiments, the molar ratio of the compound to the neutral lipid is from about 2:1 to about 8:1.
在一些实施方案中，所述化合物和所述类固醇的摩尔比为约5:1至1:1.In some embodiments, the molar ratio of the compound to the steroid is about 5:1 to 1:1.
在一些实施方案中，所述化合物和所述聚合物缀合的脂质的摩尔比为约100:1至约20:1.In some embodiments, the molar ratio of the compound to the polymer-conjugated lipid is about 100:1 to about 20:1.
在一些实施方案中，所述治疗剂包含核酸。In some embodiments, the therapeutic agent comprises a nucleic acid.
在一些实施方案中，所述治疗剂包含CRISPR系统的组分和/或编码CRISPR系统的组分的核酸。In some embodiments, the therapeutic agent comprises a component of a CRISPR system and/or a nucleic acid encoding a component of a CRISPR system.
在一些实施方案中，所述治疗剂包含guide RNA和/或编码Cas酶的核酸。In some embodiments, the therapeutic agent comprises a guide RNA and/or a nucleic acid encoding a Cas enzyme.
另一方面，本申请提供了一种细胞，其包含所述的化合物和/或所述的组合物。In another aspect, the present application provides a cell comprising the compound and/or the composition.
另一方面，本申请提供了一种试剂盒，其包含所述的化合物、所述的组合物和/或所述的细胞。On the other hand, the present application provides a kit comprising the compound, the composition and/or the cell.
另一方面，本申请提供了一种施用治疗剂的方法，其包含提供所述的化合物、所述的组合物、所述的细胞和/或所述的试剂盒，所述治疗剂与所述化合物混合，或所述治疗剂存在于所述组合物、所述细胞和/或所述试剂盒中。On the other hand, the present application provides a method for administering a therapeutic agent, which comprises providing the compound, the composition, the cell and/or the kit, wherein the therapeutic agent is mixed with the compound, or the therapeutic agent is present in the composition, the cell and/or the kit.
另一方面，本申请提供了一种递送核酸的方法，其包含提供所述的化合物、所述的组合物、所述的细胞和/或所述的试剂盒，所述核酸与所述化合物混合，或所述核酸存在于所述组合物、所述细胞和/或所述试剂盒中。 On the other hand, the present application provides a method for delivering nucleic acid, which comprises providing the compound, the composition, the cell and/or the kit, wherein the nucleic acid is mixed with the compound, or the nucleic acid is present in the composition, the cell and/or the kit.
另一方面，本申请提供了一种所述的化合物、所述的组合物、所述的细胞和/或所述的试剂盒在制备药物中的用途，所述药物用于治疗和/或预防疾病或病症的方法。On the other hand, the present application provides a use of the compound, the composition, the cell and/or the kit in the preparation of a drug, wherein the drug is used in a method for treating and/or preventing a disease or condition.
本领域技术人员能够从下文的详细描述中容易地洞察到本申请的其它方面和优势。下文的详细描述中仅显示和描述了本申请的示例性实施方式。如本领域技术人员将认识到的，本申请的内容使得本领域技术人员能够对所公开的具体实施方式进行改动而不脱离本申请所涉及发明的精神和范围。相应地，本申请的附图和说明书中的描述仅仅是示例性的，而非为限制性的。Those skilled in the art can easily perceive other aspects and advantages of the present application from the detailed description below. In the detailed description below, only exemplary embodiments of the present application are shown and described. As will be appreciated by those skilled in the art, the content of the present application enables those skilled in the art to modify the disclosed specific embodiments without departing from the spirit and scope of the invention to which the present application relates. Accordingly, the description in the drawings and specification of the present application is merely exemplary and not restrictive.
附图说明BRIEF DESCRIPTION OF THE DRAWINGS
本申请所涉及的发明的具体特征如所附权利要求书所显示。通过参考下文中详细描述的示例性实施方式和附图能够更好地理解本申请所涉及发明的特点和优势。对附图简要说明如下：The specific features of the invention involved in this application are shown in the attached claims. The features and advantages of the invention involved in this application can be better understood by referring to the exemplary embodiments and drawings described in detail below. The drawings are briefly described as follows:
图1显示的是本申请递送物质用于Luciferase在小鼠肝脏中的表达效果。FIG. 1 shows the expression effect of the delivery substance of the present application on Luciferase in mouse liver.
图2显示的是本申请递送物质用于Ai9转基因的成年小鼠静脉注射肝脏转染效果评价。FIG2 shows the evaluation of the effect of intravenous injection of the delivery material of the present application on liver transfection in adult Ai9 transgenic mice.
图3显示的是本申请递送物质用于活体内小鼠PCSK9基因编辑效果评价。FIG3 shows the evaluation of the PCSK9 gene editing effect of the delivered material of the present application in vivo in mice.
具体实施方式Detailed ways
以下由特定的具体实施例说明本申请发明的实施方式，熟悉此技术的人士可由本说明书所公开的内容容易地了解本申请发明的其他优点及效果。The following is an explanation of the implementation of the present invention by means of specific embodiments. Those skilled in the art can easily understand other advantages and effects of the present invention from the contents disclosed in this specification.
术语定义Definition of Terms
在本申请中，术语“化合物”通常指具有两种或两种以上不同元素的物质。例如，本申请的化合物可以是有机化合物，例如本申请的化合物可以是分子量500以下的化合物，可以是分子量1000以下的化合物，也可以是分子量1000以上的化合物，也可以是10000以上、100000以上的化合物。在本申请中，化合物还可以是指通过化学键相连的化合物，例如可以是一个或多个分子量1000以下的分子通过化学键与生物大分子相连的化合物，所述生物大分子可以是高聚糖、蛋白、核酸、多肽等。例如本申请的化合物可以包括蛋白质与一个或多个分子量1000以下的分子相连的化合物，可以是包括蛋白质与一个或多个分子量10000以下的分子相连的化合物，可以是包括蛋白质与一个或多个分子量100000以下的分子相连的化合物。 In the present application, the term "compound" generally refers to a substance having two or more different elements. For example, the compound of the present application may be an organic compound, for example, the compound of the present application may be a compound with a molecular weight of less than 500, a compound with a molecular weight of less than 1000, a compound with a molecular weight of more than 1000, or a compound with a molecular weight of more than 10000 or more than 100000. In the present application, a compound may also refer to a compound connected by chemical bonds, for example, a compound in which one or more molecules with a molecular weight of less than 1000 are connected to a biomacromolecule by chemical bonds, and the biomacromolecule may be a high polysaccharide, protein, nucleic acid, polypeptide, etc. For example, the compound of the present application may include a compound in which a protein is connected to one or more molecules with a molecular weight of less than 1000, a compound in which a protein is connected to one or more molecules with a molecular weight of less than 10000, or a compound in which a protein is connected to one or more molecules with a molecular weight of less than 100000.
如本文所用，术语“本申请化合物”指本申请的化合物。该术语还包括本申请化合物的各种晶型形式、药学上可接受的盐、水合物或溶剂合物。本领域技术人员还将认识到，“前药”通常是指该前药物质在对受试者施用并且之后可以代谢形成具有本申请化合物结构的物质，“代谢产物”通常是指本申请化合物在对受试者施用并且之后代谢得到的物质，这类衍生物所包含的前药和代谢产物包括在本发明的范围内。As used herein, the term "compound of the present application" refers to the compound of the present application. The term also includes various crystalline forms, pharmaceutically acceptable salts, hydrates or solvates of the compound of the present application. Those skilled in the art will also recognize that "prodrug" generally refers to the prodrug substance that can be metabolized to form a substance having the structure of the compound of the present application after being administered to a subject, and "metabolite" generally refers to the substance obtained by the metabolism of the compound of the present application after being administered to a subject, and the prodrugs and metabolites contained in such derivatives are included within the scope of the present invention.
在本申请中，本申请的化合物包含化合物的互变异构体、内消旋体、外消旋体、对映异构体、和/或非对映异构体。在本申请中，术语“非对映异构体”通常是指具有两个或更多个手性中心并且其分子不是彼此的镜像的立体异构体。非对映异构体可以具有不同的物理性质，例如、熔点、沸点、波谱性质和反应性。在本申请中，术语“互变异构体”或“互变异构形式”可互换使用，通常是指可通过低能垒(low energy barrier)互相转化的不同能量的结构异构体。例如，质子互变异构体(protontautomer)(也称为质子移变互变异构体(prototropic tautomer))包括通过质子迁移进行的互相转化，诸如酮-烯醇异构化和亚胺-烯胺异构化。价键互变异构体(valence tautomer)包括通过一些成键电子的重组进行的互相转化。在本申请中，术语“内消旋体”通常是指分子内含有不对称性的原子，但因其具有对称因素而使分子内总旋光度为零。术语“外消旋体”或“外消旋混合物”是指由等摩尔量的两种对映异构体物质构成的组合物。In the present application, the compounds of the present application include tautomers, mesomorphs, racemates, enantiomers, and/or diastereomers of the compounds. In the present application, the term "diastereomer" generally refers to a stereoisomer having two or more chiral centers and whose molecules are not mirror images of each other. Diastereomers can have different physical properties, such as melting points, boiling points, spectral properties, and reactivity. In the present application, the terms "tautomer" or "tautomeric form" are used interchangeably and generally refer to structural isomers of different energies that can be mutually converted by low energy barriers. For example, proton tautomers (also referred to as prototropic tautomers) include mutual conversions by proton migration, such as keto-enol isomerization and imine-enamine isomerization. Valence tautomers include mutual conversions by the reorganization of some bonding electrons. In this application, the term "mesomeric compound" generally refers to a molecule containing an asymmetric atom, but due to its symmetry factors, the total optical rotation of the molecule is zero. The term "racemic compound" or "racemic mixture" refers to a composition composed of equimolar amounts of two enantiomeric substances.
除非特别说明，本申请中，所有出现的化合物均意在包括所有可能的光学异构体，如单一手性的化合物，或各种不同手性化合物的混合物(即外消旋体)。本申请的所有化合物之中，各手性碳原子可以任选地为R构型或S构型，或R构型和S构型的混合物。Unless otherwise specified, in this application, all compounds appearing are intended to include all possible optical isomers, such as single chiral compounds, or mixtures of various chiral compounds (i.e., racemates). In all compounds of this application, each chiral carbon atom can optionally be in R configuration or S configuration, or a mixture of R configuration and S configuration.
在本申请中，本申请的化合物的某些原子可能以一种以上的同位素形式出现。例如，氢可能以氕(1H)、氘(2H)和氚(3H)的形式存在，碳可能以三种不同的同位素(12C、13C和14C)自然存在。可并入本申请化合物中的同位素示例还包括但不限于15N、18O、17O、18F、32P、33P、129I、131I、123I、124I、125I，或者类似的同位素。因此，相对于这些同位素的自然丰度，本申请的化合物可富集在一种或多种这些同位素中。如本领域技术人员所知，此类同位素富集化合物可用于多种用途。例如，用重同位素如氘(2H)替代可能会提供某些治疗优势，这可以是由于更高的代谢稳定性。例如，氘(2H)的自然丰度约为0.015％。因此，自然界中大约每6500个氢原子，就有一个氘原子。因此，本申请的含氘化合物在一个或多个位置(视情况而定)的氘丰度大于0.015％。除非另有指明，否则本申请所述的结构还可以包括仅在是否存在一个或多个同位素富集原子方面存在差别的化合物。举例而言，除了氢原子被氘或氚所取代，或碳原子被碳13或碳14所取代之外，其余部分均与本申请结构一致的化合物均在 本申请的范围之内。In the present application, some atoms of the compounds of the present application may appear in more than one isotopic form. For example, hydrogen may exist in the form of protium ( 1H ), deuterium ( 2H ) and tritium ( 3H ), and carbon may exist naturally in three different isotopes ( 12C , 13C and 14C ). Examples of isotopes that can be incorporated into the compounds of the present application also include but are not limited to15N , 18O , 17O , 18F , 32P , 33P , 129I , 131I , 123I , 124I , 125I , or similar isotopes. Therefore, relative to the natural abundance of these isotopes, the compounds of the present application can be enriched in one or more of these isotopes. As known to those skilled in the art, such isotope-enriched compounds can be used for a variety of purposes. For example, substitution with heavy isotopes such as deuterium ( 2H ) may provide certain therapeutic advantages, which may be due to higher metabolic stability. For example, the natural abundance of deuterium ( 2H ) is about 0.015%. Therefore, there is one deuterium atom for approximately every 6500 hydrogen atoms in nature. Therefore, the deuterium-containing compounds of the present application have a deuterium abundance greater than 0.015% at one or more positions (as the case may be). Unless otherwise indicated, the structures described in the present application may also include compounds that differ only in the presence or absence of one or more isotopically enriched atoms. For example, compounds that are consistent with the structures of the present application except that hydrogen atoms are replaced by deuterium or tritium, or carbon atoms are replaced by carbon 13 or carbon 14, are all in Within the scope of this application.
在本申请中，术语“药学上可接受的”是指在合理医学判断范围内，适用于与人类和动物组织接触而没有过量毒性、刺激、过敏反应或其它问题或并发症、与合理的利益/风险比相称的物质。在本申请中，术语“药学上可接受的盐”是指如上文所定义的药学上可接受的并且具有期望的药理学活性的所述化合物的盐，其包括所述化合物的酸加成盐和碱加成盐。其中，“药学上可接受的酸加成盐”是指保留了游离碱的生物效力和性能，不是生物学上或其他方面不期望的，并且与无机酸(例如但不限于盐酸、氢溴酸、硫酸、硝酸、磷酸等)或与有机酸(例如但不限于乙酸、2,2-二氯乙酸、己二酸、藻酸、抗坏血酸、天冬氨酸、苯磺酸、苯甲酸、4-乙酰氨基苯甲酸、樟脑酸、樟脑-10-磺酸、癸酸、己酸、辛酸、碳酸、肉桂酸、柠檬酸、环拉酸、十二烷基硫酸、乙烷-1,2-二磺酸、乙磺酸、2-羟基乙磺酸、甲酸、富马酸、半乳糖二酸、龙胆酸、葡庚糖酸、葡萄糖醛酸、谷氨酸、戊二酸、2-氧代-戊二酸、甘油磷酸、乙醇酸、马尿酸、异丁酸、乳酸、乳糖酸、月桂酸、马来酸、苹果酸、丙二酸、扁桃酸、甲磺酸、粘酸、萘-1,5-二磺酸、萘-2-磺酸、1-羟基-2-萘甲酸、烟酸、油酸、乳清酸、草酸、棕榈酸、双羟萘酸、丙酸、焦谷氨酸、丙酮酸、水杨酸、4-氨基水杨酸、癸二酸、硬脂酸、琥珀酸、酒石酸、硫氰酸、对甲苯磺酸、三氟乙酸、十一碳烯酸等)形成的盐。术语“药学上可接受的碱加成盐”是指保留了游离酸的生物效力和性能，不是生物学上或其他方面不期望的盐。这些盐是通过将无机碱或有机碱加到游离酸中来制备的。衍生自无机碱的盐包括但不限于钠盐、钾盐、锂盐、铵盐、钙盐、镁盐、铁盐、锌盐、铜盐、锰盐、铝盐等。衍生自有机碱的盐包括但不限于伯胺、仲胺和叔胺的盐，取代的胺，其包括天然存在的取代的胺、环胺和碱性离子交换树脂，如氨、异丙胺、三甲胺、二乙胺、三乙胺、三丙胺、二乙醇胺、乙醇胺、地阿诺、2-二甲基氨基乙醇、2-二乙基氨基乙醇、二环己胺、赖氨酸、精氨酸、组氨酸、咖啡因、普鲁卡因、海巴明、胆碱、甜菜碱、苯那明、苄星、乙二胺、葡糖胺、甲基葡糖胺、可可碱、三乙醇胺、氨丁三醇、嘌呤、哌嗪、哌啶、N-乙基哌啶、多胺树脂等。In the present application, the term "pharmaceutically acceptable" refers to substances that are suitable for contact with human and animal tissues without excessive toxicity, irritation, allergic reaction or other problems or complications, and are commensurate with a reasonable benefit/risk ratio, within the scope of reasonable medical judgment. In the present application, the term "pharmaceutically acceptable salt" refers to a salt of the compound that is pharmaceutically acceptable as defined above and has the desired pharmacological activity, including acid addition salts and base addition salts of the compound. The term "pharmaceutically acceptable acid addition salt" refers to a free base which retains the biological effectiveness and properties of the free base, is not biologically or otherwise undesirable, and is soluble in water and is soluble in water and is stable to water and is stable to water. The salts formed by the pharmaceutically acceptable base addition salts include maleic acid, galactaric acid, gentisic acid, glucoheptonic acid, glucuronic acid, glutamic acid, glutaric acid, 2-oxo-glutaric acid, glycerophosphoric acid, glycolic acid, hippuric acid, isobutyric acid, lactic acid, lactobionic acid, lauric acid, maleic acid, malic acid, malonic acid, mandelic acid, methanesulfonic acid, mucic acid, naphthalene-1,5-disulfonic acid, naphthalene-2-sulfonic acid, 1-hydroxy-2-naphthoic acid, nicotinic acid, oleic acid, orotic acid, oxalic acid, palmitic acid, pamoic acid, propionic acid, pyroglutamic acid, pyruvic acid, salicylic acid, 4-aminosalicylic acid, sebacic acid, stearic acid, succinic acid, tartaric acid, thiocyanic acid, p-toluenesulfonic acid, trifluoroacetic acid, undecylenic acid, etc. The term "pharmaceutically acceptable base addition salt" refers to a salt that retains the biological effectiveness and performance of the free acid and is not biologically or otherwise undesirable. These salts are prepared by adding an inorganic base or an organic base to the free acid. Salts derived from inorganic bases include, but are not limited to, sodium, potassium, lithium, ammonium, calcium, magnesium, iron, zinc, copper, manganese, aluminum, etc. Salts derived from organic bases include, but are not limited to, salts of primary, secondary, and tertiary amines, substituted amines including naturally occurring substituted amines, cyclic amines, and basic ion exchange resins such as ammonia, isopropylamine, trimethylamine, diethylamine, triethylamine, tripropylamine, diethanolamine, ethanolamine, diarnol, 2-dimethylaminoethanol, 2-diethylaminoethanol, dicyclohexylamine, lysine, arginine, histidine, caffeine, procaine, hydrazine, choline, betaine, benzylidene, benzathine, ethylenediamine, glucosamine, methylglucamine, theobromine, triethanolamine, tromethamine, purine, piperazine, piperidine, N-ethylpiperidine, polyamine resins, and the like.
在本申请中，术语“基团之间包含2至6个原子”和“链长为2至6个原子的连接基团”是指2个、3个、4个、5个或6个原子经由连续的共价键互相连接形成链状的分子片段或连接基团，其中所述的原子可以是单原子(例如但不限于碳原子、氮原子、氧原子和硫原子等)、包含在任何取代基中的中心原子(例如但不限于烷基、烯基或炔基的中心碳原子和胺基的中心氮原子等)和/或任何取代基中参与形成主链的原子(例如但不限于酯基中由单键相连的碳原子和氧原子等)。在某些实施方案中，基团之间可能包含形成环状结构的多个原子或基团之间的连接基团为环状基团，此时间隔的原子数量或链长为环状结构上与主链相连接的两个原 子之间(包含两端的原子)形成最短距离的原子数量。In the present application, the terms "groups containing 2 to 6 atoms" and "linking groups with a chain length of 2 to 6 atoms" refer to 2, 3, 4, 5 or 6 atoms connected to each other via continuous covalent bonds to form a chain-like molecular fragment or linking group, wherein the atoms may be single atoms (such as but not limited to carbon atoms, nitrogen atoms, oxygen atoms and sulfur atoms, etc.), central atoms contained in any substituent (such as but not limited to the central carbon atom of an alkyl, alkenyl or alkynyl group and the central nitrogen atom of an amine group, etc.) and/or atoms participating in the formation of the main chain in any substituent (such as but not limited to the carbon atoms and oxygen atoms connected by single bonds in an ester group, etc.). In certain embodiments, the groups may contain multiple atoms forming a ring structure or the linking group between the groups is a ring group, in which case the number of atoms or the chain length of the spacer is the number of atoms between the two atoms connected to the main chain on the ring structure. The number of atoms that form the shortest distance between the atoms (including the atoms at both ends).
在本申请中，术语“可电离脂质”是指具有至少一个可质子化或可去质子化基团的脂质，例如阳离子脂质，使得该脂质在等于或低于生理pH(例如，pH7.4)的pH下带正电荷，并且在第二pH(优选等于生理pH或高于生理pH)下为中性。本领域普通技术人员将理解，根据pH添加或去除质子是一种平衡过程，并且提到带点脂质或中性脂质是指主要物质的性质，并不要求所有脂质都以带电或中性形式存在。通常，可电离脂质的可质子化基团的pKa在约4至约7的范围内，在不同pH下，其电离反应影响LNP的表面电荷，这种电荷状态可影响对于细胞内递送核酸关键的血浆蛋白吸收、血液清除和组织分布以及形成溶核内体(endosomolytic)非双层结构的能力。In the present application, the term "ionizable lipid" refers to a lipid having at least one protonatable or deprotonatable group, such as a cationic lipid, such that the lipid is positively charged at a pH equal to or lower than physiological pH (e.g., pH 7.4) and is neutral at a second pH (preferably equal to or higher than physiological pH). It will be understood by those of ordinary skill in the art that the addition or removal of protons according to pH is an equilibrium process, and reference to charged lipids or neutral lipids refers to the properties of the primary substance and does not require that all lipids exist in a charged or neutral form. Typically, the pKa of the protonatable group of an ionizable lipid is in the range of about 4 to about 7, and at different pHs, its ionization reaction affects the surface charge of the LNP, and this charge state can affect the absorption of plasma proteins, blood clearance, and tissue distribution, which are critical for intracellular delivery of nucleic acids, as well as the ability to form endosomolytic non-bilayer structures.
在本申请中，术语“阳离子提供基团”通常是指可电离脂质中带有正电荷的头部基团，其基团大小和电荷密度对封装核酸的效率，稳定LNP，与细胞膜相互作用及促进内体逃逸等过程产生影响。常见的可电离脂质仅包含一个头部基团，有些也包含几个头部基团，非限制性示例的阳离子提供基团包括胺(伯胺、仲胺、叔胺、季胺)、胍、杂环基团等。例如，临床上使用的可电离脂质DLin-MC3-DMA、SM-102和ALC-0315包含叔胺头部，其可发生pH依赖性的阳离子化。阳离子提供基团可以是取代的或非取代的。In the present application, the term "cation providing group" generally refers to a head group with a positive charge in an ionizable lipid, and its group size and charge density have an impact on the efficiency of encapsulating nucleic acids, stabilizing LNP, interacting with cell membranes, and promoting processes such as endosome escape. Common ionizable lipids contain only one head group, and some also contain several head groups. Non-limiting examples of cation providing groups include amines (primary amines, secondary amines, tertiary amines, quaternary amines), guanidines, heterocyclic groups, etc. For example, the ionizable lipids DLin-MC3-DMA, SM-102, and ALC-0315 used clinically contain tertiary amine heads, which can undergo pH-dependent cationization. The cation providing group can be substituted or non-substituted.
在本申请中，术语“氢键提供基团”通常是指可电离脂质中能够作为氢键供体与核酸的碱基形成分子间弱相互作用力的连接基团或作为连接基团一部分的基团，非限制性示例的氢键提供基团包括脲基、酰胺、羧基、二价过渡金属等。氢键提供基团可以是取代的或非取代的。In the present application, the term "hydrogen bond providing group" generally refers to a linking group or a group as part of a linking group in an ionizable lipid that can form a weak intermolecular interaction force with a base of a nucleic acid as a hydrogen bond donor, and non-limiting examples of hydrogen bond providing groups include urea, amide, carboxyl, divalent transition metal, etc. The hydrogen bond providing group may be substituted or unsubstituted.
在本申请中，术语“尾部具有疏水性”是指具有非极性基团的尾部基团，所述非极性基团包括但不限于长链饱和和不饱和脂族烃基、以及任选地被一个或多个芳族、环脂族或杂环基团取代的此类基团。优选的实例包括但不限于二酰基甘油、二烷基甘油、N-N-二烷基氨基、1,2-二酰氧基-3-氨基丙烷和1,2-二烷基-3-氨基丙烷。In the present application, the term "the tail has hydrophobicity" refers to a tail group having a non-polar group, and the non-polar group includes, but is not limited to, long-chain saturated and unsaturated aliphatic hydrocarbon groups, and such groups optionally substituted with one or more aromatic, cycloaliphatic or heterocyclic groups. Preferred examples include, but are not limited to, diacylglycerol, dialkylglycerol, N-N-dialkylamino, 1,2-diacyloxy-3-aminopropane and 1,2-dialkyl-3-aminopropane.
在本申请中，术语“主链”在针对某个基团的情形下，通常是指含碳原子较多的碳链，或者包含官能团的最长碳链为主链。在某些实施方案中，当基团具有相同长度的链可作为主链时，选定具有支链数目最多(或支链最简单)的碳链作为其主链。In this application, the term "main chain" in the case of a certain group generally refers to a carbon chain with more carbon atoms, or the longest carbon chain containing a functional group as the main chain. In certain embodiments, when a group has chains of the same length that can be used as the main chain, the carbon chain with the largest number of branches (or the simplest branch) is selected as its main chain.
在本申请中，术语“任选取代的取代基”和“任选地被取代”是指所述参考基团可被一个或多个额外基团取代或未被取代，所述额外基团单独且独立地选自烷基、烯基、炔基、环烷基、芳基、杂芳基、杂环烷基、羟基、烷氧基、巯基、氰基、卤素、羰基、硫代羰基、异氰酸基、硫氰酸基、异硫氰酸基、硝基、全卤代烷基、全氟代烷基和包括单取代及双取代氨基 基团的氨基，及其受保护的衍生物。任选取代基的非限制性示例包括卤素、-CN、＝O、＝N-OH、＝N-OR、＝N-R、OR、-C(O)R、-C(O)OR、-OC(O)R、-OC(O)OR、-C(O)NHR、-C(O)NR2、-OC(O)NHR、-OC(O)NR2、-SR-、-S(O)R、-S(O)2R、-NHR、-N(R)2、-NHC(O)R、-NRC(O)R、-NHC(O)OR、-NRC(O)OR、S(O)2NHR、-S(O)2N(R)2、-NHS(O)2NR2、-NRS(O)2NR2、-NHS(O)2R、-NRS(O)2R、C1-C8烷基、C1-C8烷氧基、芳基、杂芳基、环烷基、杂环烷基、卤素取代的C1-C8烷基、和卤素取代的C1-C8烷氧基，其中各R独立选自H、卤素、C1-C8烷基、C1-C8烷氧基、芳基、杂芳基、环烷基、杂环烷基、卤素取代的C1-C8烷基、和卤素取代的C1-C8烷氧基。此类取代基团的位置和数量按各基团熟知的价态限制确定，例如＝O是烷基的合适取代基但不适于芳基。As used herein, the terms "optionally substituted substituents" and "optionally substituted" mean that the referenced group may be substituted or unsubstituted by one or more additional groups individually and independently selected from the group consisting of alkyl, alkenyl, alkynyl, cycloalkyl, aryl, heteroaryl, heterocycloalkyl, hydroxy, alkoxy, mercapto, cyano, halogen, carbonyl, thiocarbonyl, isocyanato, thiocyanato, isothiocyanato, nitro, perhaloalkyl, perfluoroalkyl and mono- and di-substituted amino groups. The amino group and its protected derivatives. Non-limiting examples of optional substituents include halogen, -CN, =O, =N-OH, =N-OR, =NR, OR, -C(O)R, -C(O)OR, -OC(O)R, -OC(O)OR, -C(O)NHR, -C(O) NR2 , -OC(O)NHR, -OC(O) NR2 , -SR-, -S(O)R, -S(O) 2R , -NHR, -N(R) 2 , -NHC(O)R, -NRC(O)R, -NHC(O)OR, -NRC(O)OR, S(O) 2NHR , -S(O ) 2N (R) 2 , -NHS(O) 2NR2 , -NRS(O) 2NR2 , -NHS(O) 2R , -NRS(O) 2R , C1 - C8 alkyl, C1-C8 alkyl, C1- C8 C 1 -C 8 alkoxy, aryl, heteroaryl, cycloalkyl, heterocycloalkyl, halogen-substituted C 1 -C 8 alkyl, and halogen-substituted C 1 -C 8 alkoxy, wherein each R is independently selected from H, halogen, C 1 -C 8 alkyl, C 1 -C 8 alkoxy, aryl, heteroaryl, cycloalkyl, heterocycloalkyl, halogen-substituted C 1 -C 8 alkyl, and halogen-substituted C 1 -C 8 alkoxy. The position and number of such substituent groups are determined by the well-known valence restrictions of each group, for example, =O is a suitable substituent for alkyl but not for aryl.
在本申请中，术语“烷基”通常是指烷除去氢原子所衍生的残基。烷基可以是取代的或非取代的，替代或者非替代的。术语“烷基”通常指饱和的直链或支链脂肪族烃基，其具有从母体烷的相同碳原子或两个不同的碳原子上除去氢原子所衍生的残基，其可以为包含1至20个碳原子的直链或支链基团，例如含有1至12个碳原子，例如含有1至6个碳原子的链烷基。烷基的非限制性实例包括但不限于甲基、乙基、丙基、丙基、丁基等。烷基可以是取代的或非取代的，替代或者非替代的，例如当被取代时，取代基可以在任何可使用的连接点上被取代，所述取代基可以独立地任选选自烷基、烯基、炔基、烷氧基、烷硫基、烷基氨基、卤素、巯基、羟基、硝基、氰基、环烷基、杂环基、芳基、杂芳基、环烷氧基、杂环烷氧基、环烷硫基、杂环烷硫基和氧代基中的一个或多个取代基所取代，例如可以是氢、氕、氘、氚、卤素、-NO2、-CN、-OH、-SH、-NH2、-C(O)H、-CO2H、-C(O)C(O)H、-C(O)CH2C(O)H、-S(O)H、-S(O)2H、-C(O)NH2、-SO2NH2、-OC(O)H、-N(H)SO2H或C1-6脂肪族基团。In the present application, the term "alkyl" generally refers to an alkane residue derived from a hydrogen atom. Alkyl can be substituted or non-substituted, substituted or non-substituted. The term "alkyl" generally refers to a saturated straight or branched aliphatic hydrocarbon radical, which has a residue derived from a hydrogen atom removed from the same carbon atom or two different carbon atoms of the parent alkane, which can be a straight or branched group containing 1 to 20 carbon atoms, for example, containing 1 to 12 carbon atoms, for example, a chain alkyl containing 1 to 6 carbon atoms. Non-limiting examples of alkyl include, but are not limited to, methyl, ethyl, propyl, propyl, butyl, etc. The alkyl group may be substituted or unsubstituted, substituted or unsubstituted, for example, when substituted, the substituent may be substituted at any available point of attachment, and the substituent may be independently selected from one or more substituents selected from alkyl, alkenyl, alkynyl, alkoxy, alkylthio, alkylamino, halogen, mercapto, hydroxy, nitro, cyano, cycloalkyl, heterocyclyl, aryl, heteroaryl, cycloalkyloxy, heterocycloalkyloxy, cycloalkylthio, heterocycloalkylthio and oxo, for example, hydrogen, protium, deuterium, tritium, halogen, -NO2 , -CN, -OH, -SH, -NH2 , -C(O)H, -CO2H , -C(O)C(O)H, -C(O) CH2C (O)H, -S(O)H, -S(O) 2H , -C(O) NH2 , -SO2NH2 , -OC(O)H , -N(H) SO2H or C 1-6 aliphatic groups.
在本申请中，术语“烯基”通常是指含有一个或多个双键的直链或支链烃基。烯基的示例性实例包括烯丙基、高烯丙基、乙烯基、巴豆基、丁烯基、戊烯基和己烯基等。具有一个以上双键的C2-6链烯基的示例性实例包括丁二烯基、戊二烯基、己二烯基和己三烯基以及它们的支化形式。不饱和键(双键)的位置可以是在碳链的任何一个位置。烯基可以是取代的或非取代的。In the present application, the term "alkenyl" generally refers to a straight or branched hydrocarbon group containing one or more double bonds. Illustrative examples of alkenyl include allyl, homoallyl, vinyl, crotyl, butenyl, pentenyl and hexenyl, etc. Illustrative examples of C2-6 alkenyl with more than one double bond include butadienyl, pentadienyl, hexadienyl and hexatrienyl and their branched forms. The position of the unsaturated bond (double bond) can be at any position of the carbon chain. Alkenyl can be substituted or unsubstituted.
在本申请中，术语“炔基”通常是指不饱和直链或支链炔基，例如乙炔基、1-丙炔基、炔丙基、丁炔基等。炔基可以是取代的或非取代的。In the present application, the term "alkynyl" generally refers to an unsaturated straight or branched chain alkynyl group, for example, ethynyl, 1-propynyl, propargyl, butynyl, etc. The alkynyl group may be substituted or unsubstituted.
在本申请中，术语“烃基”通常是指仅由碳原子和氢原子组成的直链或支链或环状烃原子团或其组合，它可以是完全饱和的、单不饱和或多不饱和的，可以包括二价和多价原子团，具有所指定的碳原子数(也就是说C1-C10表示一至十个碳)。饱和烃原子团的实例包括但不限 于甲基、乙基、正丙基、异丙基、正丁基、叔丁基、异丁基、仲丁基、环己基、(环己基)甲基、环丙基甲基、正戊基、正己基、正庚基、正辛基的同系物和异构体等。不饱和的烃基是具有一条或多条双键或叁键者。不饱和的烃基实例包括但不限于乙烯基、2-丙烯基、巴豆基、2-异戊烯基、2-(丁二烯基)、2,4-戊二烯基、3-(1,4-戊二烯基)、乙炔基、1-与3-丙炔基、3-丁炔基和高级同系物和异构体。术语“烃基”除非有相反注解，还意味着包括下列详细定义的那些烃基衍生物，例如“杂烃基”。限于烃类基团的烃基也称“均烃基”。In this application, the term "hydrocarbyl" generally refers to a straight or branched chain or cyclic hydrocarbon radical or combinations thereof consisting only of carbon atoms and hydrogen atoms, which may be fully saturated, monounsaturated or polyunsaturated, and may include divalent and polyvalent radicals, having a specified number of carbon atoms (i.e., C1 - C10 means one to ten carbons). Examples of saturated hydrocarbon radicals include, but are not limited to Homologs and isomers of methyl, ethyl, n-propyl, isopropyl, n-butyl, tert-butyl, isobutyl, sec-butyl, cyclohexyl, (cyclohexyl)methyl, cyclopropylmethyl, n-pentyl, n-hexyl, n-heptyl, n-octyl, etc. Unsaturated hydrocarbon groups are those having one or more double bonds or triple bonds. Examples of unsaturated hydrocarbon groups include, but are not limited to, vinyl, 2-propenyl, crotyl, 2-isopentenyl, 2-(butadienyl), 2,4-pentadienyl, 3-(1,4-pentadienyl), ethynyl, 1- and 3-propynyl, 3-butynyl, and higher homologs and isomers. The term "hydrocarbon group" is also meant to include those hydrocarbon derivatives defined in detail below, such as "heterohydrocarbon groups", unless otherwise noted. Hydrocarbon groups limited to hydrocarbon groups are also referred to as "homohydrocarbon groups".
在本申请中，术语“脂环基”通常是指具有从脂肪环的相同碳原子或多个不同的碳原子上除去氢原子所衍生的残基。术语“环烷”通常指饱和或部分不饱和单环或多环环状烃，碳环包含3至20个碳原子，可以包含3至12个碳原子，可以包含3至10个碳原子，可以包含3至8个碳原子。脂环基的非限制性实例包括环丙烷基、环丁烷基、环戊烷基、环戊烯基、环己烷基、环己烯基、环己二烯基、环庚烷基、环庚三烯基、环辛烷基等；多环碳环可以包括螺环、稠环和桥环的碳环。脂环基可以是取代的或非取代的。在本申请中，术语“碳环基”通常是指具有碳环的碳原子上除去一个氢原子所衍生的残基。术语“碳环”通常指饱和或部分不饱和单环或多环环状烃，碳环包含3至20个碳原子，可以包含3至12个碳原子，可以包含3至10个碳原子，可以包含3至8个碳原子。单环碳环的非限制性实例包括环丙烷、环丁烷、环戊烷、环戊烯、环己烷、环己烯、环己二烯、环庚烷、环庚三烯、环辛烷等；多环碳环可以包括螺环、稠环和桥环的碳环。碳环基可以是取代的或非取代的。在某些情形下，脂环和碳环可以相互替代使用。In the present application, the term "alicyclic group" generally refers to a residue derived from the same carbon atom or multiple different carbon atoms of an aliphatic ring by removing a hydrogen atom. The term "cycloalkane" generally refers to a saturated or partially unsaturated monocyclic or polycyclic cyclic hydrocarbon, and the carbocyclic ring contains 3 to 20 carbon atoms, can contain 3 to 12 carbon atoms, can contain 3 to 10 carbon atoms, and can contain 3 to 8 carbon atoms. Non-limiting examples of alicyclic groups include cyclopropane, cyclobutane, cyclopentane, cyclopentenyl, cyclohexane, cyclohexenyl, cyclohexadienyl, cycloheptane, cycloheptatrienyl, cyclooctane, etc.; polycyclic carbocyclic rings can include spirocyclic, condensed and bridged carbocyclic rings. Alicyclic groups can be substituted or unsubstituted. In the present application, the term "carbocyclic group" generally refers to a residue derived from the removal of a hydrogen atom from a carbon atom of a carbocyclic ring. The term "carbocycle" generally refers to a saturated or partially unsaturated monocyclic or polycyclic cyclic hydrocarbon, the carbocycle containing 3 to 20 carbon atoms, can contain 3 to 12 carbon atoms, can contain 3 to 10 carbon atoms, can contain 3 to 8 carbon atoms. Non-limiting examples of monocyclic carbocycles include cyclopropane, cyclobutane, cyclopentane, cyclopentene, cyclohexane, cyclohexene, cyclohexadiene, cycloheptane, cycloheptatriene, cyclooctane, etc.; polycyclic carbocycles can include spirocyclic, condensed and bridged carbocycles. Carbocyclic groups can be substituted or unsubstituted. In some cases, alicyclic and carbocyclic rings can be used interchangeably.
在本申请中，术语“部分不饱和的”通常是指环状结构中环分子间至少含一个双键或三键。术语“部分不饱和”涵盖带有多处不饱和的环状结构，但并非意在包括本申请所定义的芳环或杂芳环。术语“不饱和的”表示部分具有一个或多个不饱和度。In this application, the term "partially unsaturated" generally refers to a ring structure containing at least one double bond or triple bond between the ring molecules. The term "partially unsaturated" encompasses ring structures with multiple unsaturations, but is not intended to include aromatic or heteroaromatic rings as defined in this application. The term "unsaturated" means that the moiety has one or more degrees of unsaturation.
在本申请中，术语“脂杂环基”通常是指稳定的不具有芳香性的3元-7元单环碳环结构，融合的7元-10元双环杂环结构或桥联的6元-10元双环杂环结构，这些环状结构即可以是饱和的，也可以是部分饱和的，除碳原子外，这些环状结构中还含有一个或多个杂原子，其中杂原子可以选自以下组：氧、硫和氮。例如是含有1-4个上述定义的杂原子。当用来表示脂杂环环状结构上的原子时，术语“氮”可以包括发生过取代反应的氮。例如，脂杂环基可以包含“杂环烷基”，杂环烷基可以指稳定的不具有芳香性的3元-7元单环烷结构，融合的7元-10元双环杂环结构或桥联的6元-10元双环杂环结构，除碳原子外，这些环状结构中还含有一个或多个杂原子，其中杂原子可以选自以下组：氧、硫和氮。例如是含有1-4个上述定义的杂原子。杂环烷基可以是取代的或非取代的。脂杂环基可以是取代的或非取代的。 In the present application, the term "alicyclic group" generally refers to a stable 3-7-membered monocyclic carbon ring structure without aromaticity, a fused 7-10-membered bicyclic heterocyclic structure or a bridged 6-10-membered bicyclic heterocyclic structure, which ring structures can be saturated or partially saturated. In addition to carbon atoms, these ring structures also contain one or more heteroatoms, wherein the heteroatoms can be selected from the following groups: oxygen, sulfur and nitrogen. For example, it contains 1-4 heteroatoms as defined above. When used to represent atoms on the alicyclic heterocyclic ring structure, the term "nitrogen" can include nitrogen that has undergone substitution reactions. For example, the alicyclic group can include "heterocycloalkyl", which can refer to a stable 3-7-membered monocyclic alkane structure without aromaticity, a fused 7-10-membered bicyclic heterocyclic structure or a bridged 6-10-membered bicyclic heterocyclic structure, which can contain one or more heteroatoms in addition to carbon atoms, wherein the heteroatoms can be selected from the following groups: oxygen, sulfur and nitrogen. For example, it contains 1-4 heteroatoms as defined above. Heterocycloalkyl groups may be substituted or unsubstituted. Alicyclic heterocyclic groups may be substituted or unsubstituted.
在本申请中，术语“芳基”通常是指具有芳环上除去一个氢原子所衍生的残基。术语“芳环”可以指具有共轭的π电子体系的6至14元全碳单环或稠合多环(也就是共享毗邻碳原子对的环)，可以为6至10元，例如苯和萘。所述芳环可以稠合于杂芳基、杂环基或环烷基环上，其中与母体结构连接在一起的环为芳基环。芳基可以是取代的或非取代的，当被取代时，取代基可以为一个或多个以下基团，其独立地选自以下组：烷基、烯基、炔基、烷氧基、烷硫基、烷基氨基、卤素、巯基、羟基、硝基、氰基、环烷基、杂环烷基、芳基、杂芳基、环烷氧基、杂环烷氧基、环烷硫基、和杂环烷硫基。芳基可以是取代的或非取代的。In the present application, the term "aryl" generally refers to a residue derived from an aromatic ring by removing a hydrogen atom. The term "aromatic ring" may refer to a 6 to 14-membered all-carbon monocyclic ring or a fused polycyclic ring (i.e., a ring sharing adjacent carbon atom pairs) with a conjugated π electron system, which may be 6 to 10 members, such as benzene and naphthalene. The aromatic ring may be fused to a heteroaryl, heterocyclic or cycloalkyl ring, wherein the ring connected to the parent structure is an aryl ring. The aryl group may be substituted or unsubstituted, and when substituted, the substituent may be one or more of the following groups, independently selected from the following groups: alkyl, alkenyl, alkynyl, alkoxy, alkylthio, alkylamino, halogen, sulfhydryl, hydroxyl, nitro, cyano, cycloalkyl, heterocycloalkyl, aryl, heteroaryl, cycloalkyloxy, heterocycloalkyloxy, cycloalkylthio, and heterocycloalkylthio. The aryl group may be substituted or unsubstituted.
在本申请中，术语“杂芳基”通常是指具有从杂芳环的碳原子上除去一个氢原子所衍生的残基。术语“杂芳环”指包含1至4个杂原子、5至14个环原子的杂芳族体系，其中杂原子可以选自以下组：氧、硫和氮。杂芳基可以为5至10元，可以为5元或6元，例如呋喃基、噻吩基、吡啶基、吡咯基、N-烷基吡咯基、嘧啶基、吡嗪基、咪唑基、四唑基等。所述杂芳基环可以稠合于芳基、杂环基或环烷基环上，其中与母体结构连接在一起的环为杂芳基环。杂芳基可以是任选取代的或非取代的，当被取代时，取代基可以为一个或多个以下基团，其独立地选自以下组：烷基、烯基、炔基、烷氧基、烷硫基、烷基氨基、卤素、巯基、羟基、硝基、氰基、环烷基、杂环烷基、芳基、杂芳基、环烷氧基、杂环烷氧基、环烷硫基、和杂环烷硫基。杂芳基可以是取代的或非取代的。In the present application, the term "heteroaryl" generally refers to a residue derived from a carbon atom of a heteroaromatic ring by removing a hydrogen atom. The term "heteroaromatic ring" refers to a heteroaromatic system containing 1 to 4 heteroatoms, 5 to 14 ring atoms, wherein the heteroatoms can be selected from the following groups: oxygen, sulfur and nitrogen. The heteroaryl group can be 5 to 10 yuan, and can be 5 yuan or 6 yuan, such as furanyl, thienyl, pyridyl, pyrrolyl, N-alkylpyrrolyl, pyrimidyl, pyrazinyl, imidazolyl, tetrazolyl, etc. The heteroaryl ring can be fused to an aryl, heterocyclyl or cycloalkyl ring, wherein the ring connected to the parent structure is a heteroaryl ring. Heteroaryl can be optionally substituted or unsubstituted, and when substituted, the substituent can be one or more of the following groups, which are independently selected from the following groups: alkyl, alkenyl, alkynyl, alkoxy, alkylthio, alkylamino, halogen, sulfhydryl, hydroxyl, nitro, cyano, cycloalkyl, heterocycloalkyl, aryl, heteroaryl, cycloalkyloxy, heterocycloalkyloxy, cycloalkylthio, and heterocycloalkylthio. Heteroaryl can be substituted or unsubstituted.
在本申请中，如本领域技术人员可知的，“烷基”、“烯基”、“环烷基”等之类的术语可以在名称前加一个标识表示在特定情况下基团中存在的原子数，例如，C1-C4烷基，C3-C7环烷氧基，C1-C4烷基羰基氨基等，“C”后所跟下标数字表示在基团中存在的碳原子数。例如，C3烷基是指具有三个碳原子的烷基(例如，正丙基，异丙基)；C1-10中，基团的成员可具有落入1-10范围内的任何数目的碳原子。In this application, as known to those skilled in the art, terms such as "alkyl", "alkenyl", "cycloalkyl", etc. may be preceded by an identifier to indicate the number of atoms present in the group in a particular case, for example, C 1 -C 4 alkyl, C 3 -C 7 cycloalkyloxy, C 1 -C 4 alkylcarbonylamino, etc., and the subscript number following "C" indicates the number of carbon atoms present in the group. For example, C 3 alkyl refers to an alkyl group having three carbon atoms (e.g., n-propyl, isopropyl); in C 1-10 , the members of the group may have any number of carbon atoms falling within the range of 1-10.
基团中的一个或多个氢原子，例如为最多5个，例如为1-3个氢原子彼此独立地被相应数目的取代基取代。取代基仅处在它们的可能的化学位置，本领域技术人员能够在不付出过多努力的情况下确定(通过实验或理论)可能或不可能的取代。例如，具有游离氢的氨基或羟基与具有不饱和(如烯属)键的碳原子结合时可能是不稳定的。One or more hydrogen atoms in the group, for example, up to 5, for example, 1-3 hydrogen atoms are replaced by the corresponding number of substituents independently of each other. The substituents are only in their possible chemical positions, and those skilled in the art can determine (by experiment or theory) possible or impossible replacements without paying too much effort. For example, amino or hydroxyl groups with free hydrogen may be unstable when combined with carbon atoms with unsaturated (such as olefinic) bonds.
在本申请中，术语“中性脂质”是指在选定的pH下以不带电或中性的两性离子形式存在的多种脂质物质中的任一种。在生理pH下，此类脂质包括但不限于：磷脂酰胆碱如1,2-二硬脂酰基-sn-甘油基-3-磷酸胆碱(DSPC)、1,2-二棕榈酰基-sn-甘油基-3-磷酸胆碱(DPPC)、1,2-二肉豆蔻酰基-sn-甘油基-3-磷酸胆碱(DMPC)、1-棕榈酰基-2-油酰基-sn-甘油基-3-磷酸胆碱(POPC)、1,2-二油酰基-sn-甘油基-3-磷酸胆碱(DOPC)，磷脂酰乙醇胺如1,2-二油酰基-sn- 甘油基-3-磷酸乙醇胺(DOPE)，鞘磷脂(SM)，神经酰胺，类固醇如固醇及其衍生物。中性脂质可以是合成的或天然来源的。其中，术语“类固醇”是包含以下碳骨架的化合物：In the present application, the term "neutral lipid" refers to any of a variety of lipid substances that exist in an uncharged or neutral zwitterionic form at a selected pH. At physiological pH, such lipids include, but are not limited to, phosphatidylcholines such as 1,2-distearoyl-sn-glycero-3-phosphocholine (DSPC), 1,2-dipalmitoyl-sn-glycero-3-phosphocholine (DPPC), 1,2-dimyristoyl-sn-glycero-3-phosphocholine (DMPC), 1-palmitoyl-2-oleoyl-sn-glycero-3-phosphocholine (POPC), 1,2-dioleoyl-sn-glycero-3-phosphocholine (DOPC), phosphatidylethanolamines such as 1,2-dioleoyl-sn- Glyceryl-3-phosphoethanolamine (DOPE), sphingomyelin (SM), ceramides, steroids such as sterols and their derivatives. Neutral lipids can be synthetic or naturally derived. Among them, the term "steroid" is a compound containing the following carbon skeleton:
其非限制实例包括胆固醇等。 Non-limiting examples thereof include cholesterol and the like.
在本申请中，术语“聚合物缀合的脂质”是指包含脂质部分和聚合物部分的分子。聚合物缀合的脂质的实例是聚乙二醇化脂质。术语“聚乙二醇化脂质”是指包含脂质部分和聚乙二醇部分的分子。聚乙二醇化脂质在本领域中是已知的，并且包括1-(单甲氧基-聚乙二醇)-2,3-二肉豆蔻酰甘油(PEG-DMG)等。In the present application, the term "polymer-conjugated lipid" refers to a molecule comprising a lipid portion and a polymer portion. An example of a polymer-conjugated lipid is a pegylated lipid. The term "pegylated lipid" refers to a molecule comprising a lipid portion and a polyethylene glycol portion. Pegylated lipids are known in the art and include 1-(monomethoxy-polyethylene glycol)-2,3-dimyristoylglycerol (PEG-DMG) and the like.
在本申请中，术语“脂质体”通常是指通过一个或多个双层、多层、任何层状或固体的膜与外部介质隔离的具有内部空间的囊泡。例如，所述双层的膜可以通过两性分子形成，如包含空间隔离的亲水性和疏水性结构域的合成或天然来源的脂质；又例如，所述双层的膜可以通过两亲性聚合物和表面活性剂形成。脂质体包裹着水相，水相一般含有准备递送给细胞的物质，例如包含核酸。如本申请所使用的术语“脂质体”还涵盖基于脂质和聚合物的纳米颗粒。In the present application, the term "liposome" generally refers to a vesicle with an internal space that is isolated from the external medium by one or more bilayer, multilayer, any lamellar or solid membranes. For example, the bilayer membrane can be formed by amphiphilic molecules, such as synthetic or natural lipids containing spatially isolated hydrophilic and hydrophobic domains; for another example, the bilayer membrane can be formed by amphiphilic polymers and surfactants. Liposomes encapsulate an aqueous phase, which generally contains substances to be delivered to cells, such as nucleic acids. The term "liposome" as used in the present application also covers lipid- and polymer-based nanoparticles.
在本申请中，术语“核酸”是指呈单链或双链形式的含有至少两种脱氧核糖核苷酸或核糖核苷酸的聚合物，并且包括DNA、RNA及其杂交物。DNA可以呈反义分子、质粒DNA、cDNA、PCR产物或载体的形式。RNA可以呈小发卡RNA(shRNA)、信使RNA(mRNA)、反义RNA、miRNA、micRNA、多价RNA、Dicer底物RNA或病毒RNA(vRNA)及其组合的形式。核酸包括含有已知核苷酸类似物或者修饰的主链残基或连键的核酸，其是合成的、天然存在的和非天然存在的并且具有与参考核酸相似的结合特性。此类类似物的实例包括但不限于硫代磷酸酯、氨基磷酸酯、甲基磷酸酯、手性-甲基磷酸酯，2’-0-甲基核糖核苷酸和肽-核酸(PNA)。除非特别限定，否则该术语涵盖与参考核酸具有相似结合特性的含有天然核苷酸的已知类似物的核酸。除非另有说明，否则特定的核酸序列还隐含地涵盖其保守修饰的变体(如，简并密码子取代)、等位基因、直向同源物、单核苷酸多态性和互补序列以及明确指出的序列。“核苷酸”含有以下糖：脱氧核糖(DNA)或核糖(RNA)；碱基和磷酸基团。核苷酸通过磷酸基团连接在一起。“碱基”包括嘌呤和嘧啶，其进一步包括天然化合物腺嘌呤、胸腺嘧啶、鸟嘌呤、胞嘧啶、尿嘧啶、肌苷和天然类似物，以及嘌呤和嘧啶的合成衍生物， 包括但不限于放置新的反应性基团(如但不限于胺、醇、硫醇、羧化物和烃基卤化物)的修饰。In the present application, the term "nucleic acid" refers to a polymer containing at least two deoxyribonucleotides or ribonucleotides in a single-stranded or double-stranded form, and includes DNA, RNA and hybrids thereof. DNA can be in the form of antisense molecules, plasmid DNA, cDNA, PCR products or vectors. RNA can be in the form of small hairpin RNA (shRNA), messenger RNA (mRNA), antisense RNA, miRNA, micRNA, multivalent RNA, Dicer substrate RNA or viral RNA (vRNA) and combinations thereof. Nucleic acid includes nucleic acids containing known nucleotide analogs or modified main chain residues or bonds, which are synthetic, naturally occurring and non-naturally occurring and have binding properties similar to reference nucleic acids. Examples of such analogs include, but are not limited to, phosphorothioates, phosphoramidates, methyl phosphonates, chiral-methyl phosphonates, 2'-0-methyl ribonucleotides and peptide-nucleic acids (PNA). Unless otherwise specified, the term encompasses nucleic acids containing known analogs of natural nucleotides that have similar binding properties to reference nucleic acids. Unless otherwise indicated, a particular nucleic acid sequence also implicitly encompasses conservatively modified variants thereof (e.g., degenerate codon substitutions), alleles, orthologs, single nucleotide polymorphisms, and complementary sequences as well as the sequence explicitly indicated. A "nucleotide" contains the following sugars: deoxyribose (DNA) or ribose (RNA); a base and a phosphate group. The nucleotides are linked together by the phosphate groups. "Bases" include purines and pyrimidines, which further include the natural compounds adenine, thymine, guanine, cytosine, uracil, inosine, and natural analogs, as well as synthetic derivatives of purines and pyrimidines, Modifications include, but are not limited to, placement of new reactive groups such as, but not limited to, amines, alcohols, thiols, carboxylates, and hydrocarbyl halides.
在本申请中，术语“施用”是指将组合物或药剂(例如，核酸)引入个体中并且包括同时和依序引入一种或多种组合物或药剂。“施用”可以指例如治疗、药物动力学、诊断、研究、安慰剂以及实验方法。“施用”还涵盖体外和离体治疗。将组合物或药剂引入个体中是通过任何适合的途径，包括经口、经肺、经鼻、肠胃外(静脉内、肌肉内、腹膜内或皮下)、经直肠、淋巴内、瘤内或局部。施用包含自我施用和由另一个人施用。可以通过任何适合的途径进行施用。适合的施用途径使得组合物或药剂执行其预期功能。举例来说，如果所适合的途径是静脉内，那么通过将组合物或药剂引入个体的静脉来施用组合物。In the present application, the term "administering" refers to introducing a composition or medicament (e.g., nucleic acid) into an individual and includes introducing one or more compositions or medicaments simultaneously and sequentially. "Administering" may refer to, for example, treatment, pharmacokinetics, diagnosis, research, placebo, and experimental methods. "Administering" also encompasses in vitro and ex vivo treatments. Introducing a composition or medicament into an individual is by any suitable route, including oral, pulmonary, nasal, parenteral (intravenous, intramuscular, intraperitoneal or subcutaneous), rectal, intralymphatic, intratumoral or local. Administration includes self-administration and administration by another person. Administration may be performed by any suitable route. Suitable administration routes allow a composition or medicament to perform its intended function. For example, if the suitable route is intravenous, then administering the composition by introducing the composition or medicament into the vein of an individual.
在本申请中，术语“递送”是指向目标提供实体。例如，向受试者递送治疗剂和/或预防剂，可涉及对受试者施用包括治疗剂和/或预防剂的纳米颗粒组合物(例如通过静脉内、肌内、皮内或皮下途径)。对哺乳动物或哺乳动物细胞施用纳米颗粒组合物可涉及使一种或多种细胞与纳米颗粒组合物接触。其中，“接触”是指两个或更多个实体之间建立物理连接，使细胞与外部实体进行体内和离体接触的方法是生物学领域中熟知的。In this application, the term "delivery" refers to providing an entity to a target. For example, delivering a therapeutic and/or prophylactic agent to a subject may involve administering a nanoparticle composition comprising a therapeutic and/or prophylactic agent to the subject (e.g., by intravenous, intramuscular, intradermal or subcutaneous routes). Administering a nanoparticle composition to a mammal or mammalian cell may involve contacting one or more cells with the nanoparticle composition. Wherein, "contact" refers to establishing a physical connection between two or more entities, and methods for contacting a cell with an external entity in vivo and in vitro are well known in the biological field.
在本申请中，术语“预防”涵盖下列情形：(1)抑制受试者或患者的疾病发作，所述受试者或患者可能有患所述疾病的风险和/或易患所述疾病，但尚未经历或表现出所述疾病的任何或所有病状或征状；和/或(2)减慢受试者或患者中的疾病的病状或征状发作，所述受试者或患者可能有患所述疾病的风险和/或易患所述疾病，但尚未经历或表现出所述疾病的任何或所有病状或征状。As used herein, the term "prevent" or "preventing" encompasses the following situations: (1) inhibiting the onset of a disease in a subject or patient who may be at risk for and/or susceptible to the disease but who does not yet experience or display any or all of the symptoms or signs of the disease; and/or (2) slowing the onset of symptoms or signs of a disease in a subject or patient who may be at risk for and/or susceptible to the disease but who does not yet experience or display any or all of the symptoms or signs of the disease.
在本申请中，术语“治疗”涵盖下列情形：(1)抑制正经历或表现出疾病的病状或征状的受试者或患者中的所述疾病(例如，阻止所述病状和/或征状的进一步发展)，(2)改善正经历或表现出疾病的病状或征状的受试者或患者中的所述疾病(例如，逆转所述病状和/或征状)，和/或(3)实现正经历或表现出疾病的病状或征状的受试者或患者中的所述疾病的任何可测量的减轻。As used herein, the term "treating" encompasses: (1) inhibiting a disease in a subject or patient who is experiencing or exhibiting symptoms or signs of the disease (e.g., arresting further development of the symptoms and/or signs), (2) ameliorating a disease in a subject or patient who is experiencing or exhibiting symptoms or signs of the disease (e.g., reversing the symptoms and/or signs), and/or (3) achieving any measurable reduction in a disease in a subject or patient who is experiencing or exhibiting symptoms or signs of the disease.
在本申请中，术语“受试者”或“患者”是指可以施用根据本申请所述的化合物和/或组合物的任何生物体，例如用于实验、诊断、预防和/或治疗目的，典型的受试者包括动物(例如哺乳动物，如小鼠、大鼠、兔、非人灵长类动物和人)和/或植物。In the present application, the term "subject" or "patient" refers to any organism to which the compounds and/or compositions described herein can be administered, for example, for experimental, diagnostic, preventive and/or therapeutic purposes, and typical subjects include animals (e.g., mammals such as mice, rats, rabbits, non-human primates and humans) and/or plants.
在本申请中，术语“包含”通常是指包括明确指定的特征，但不排除其他要素。术语“以上”、“以下”通常是指包含本数的情况。In the present application, the term "comprising" generally refers to including the features explicitly specified, but not excluding other elements. The terms "above" and "below" generally refer to the case where the number is inclusive.
在本申请中，术语“约”通常是指在指定数值以上或以下0.5％-10％的范围内变动，例如在指定数值以上或以下0.5％、1％、1.5％、2％、2.5％、3％、3.5％、4％、4.5％、5％、5.5％、 6％、6.5％、7％、7.5％、8％、8.5％、9％、9.5％、或10％的范围内变动。In this application, the term "about" generally refers to a variation within the range of 0.5%-10% above or below the specified value, for example, 0.5%, 1%, 1.5%, 2%, 2.5%, 3%, 3.5%, 4%, 4.5%, 5%, 5.5%, 6%, 7%, 8%, 9%, 10%, 11%, 12%, 13%, 14%, 15%, 16%, 17%, 18%, 19%, 20%, 21%, 22%, 23%, 24%, 25%, 26%, 27%, 28%, 29%, 30%, 31%, 32%, 33%, 34%, 35%, 36%, 37%, 38%, 39%, 40%, 41%, 42%, 43%, 44%, 45%, 46%, 47%, 48%, 49%, 50%, 51%, 52%, 53%, 54%, 55%, 56%, 57%, 58%, 59%, 60%, 61%, 62%, 63%, 64%, 65%, 66%, 67%, 68%, 69%, 70%, 71%, 72%, 73%, 74%, 75%, 76%, 77%, 78%, 79%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 100%, 101%, 102%, 113%, 114%, 115%, 116%, 117%, 118%, 119%, 12 The price/performance ratio may vary within the range of 6%, 6.5%, 7%, 7.5%, 8%, 8.5%, 9%, 9.5%, or 10%.
发明详述DETAILED DESCRIPTION OF THE INVENTION
化合物Compound
一方面，本申请提供了一种化合物，或其药学上可接受的盐，所述化合物包含阳离子提供基团以及氢键提供基团，其所述阳离子提供基团与氢键提供基团之间包含2至6个原子，例如，所述化合物的阳离子提供基团与氢键提供基团之间包含2个、3个、4个、5个、或6个原子。On the one hand, the present application provides a compound, or a pharmaceutically acceptable salt thereof, comprising a cation providing group and a hydrogen bond providing group, wherein 2 to 6 atoms are contained between the cation providing group and the hydrogen bond providing group, for example, 2, 3, 4, 5, or 6 atoms are contained between the cation providing group and the hydrogen bond providing group of the compound.
在本申请中，所述化合物包含阳离子脂质。In the present application, the compound comprises a cationic lipid.
在本申请中，所述化合物包含可电离脂质。In the present application, the compound comprises an ionizable lipid.
在本申请中，所述阳离子提供基团包含在pH为7.0约或更低的条件下带正电的基团。例如，所述阳离子提供基团为在以下pH条件下带正电的基团：约7.0，约6.0，约5.0，约4.0，约3.0，约2.0，约1.0，约0.9，约0.8，约0.7，约0.6，约0.5，约0.4，约0.2，约0.1。In the present application, the cation providing group includes a group that is positively charged at a pH of about 7.0 or lower. For example, the cation providing group is a group that is positively charged at the following pH conditions: about 7.0, about 6.0, about 5.0, about 4.0, about 3.0, about 2.0, about 1.0, about 0.9, about 0.8, about 0.7, about 0.6, about 0.5, about 0.4, about 0.2, about 0.1.
在本申请中，所述阳离子提供基团包含在pH为4.0约或更低的条件下带正电的基团。例如，所述阳离子提供基团为在以下pH条件下带正电的基团：约4.0，约3.0，约2.0，约1.0，约0.9，约0.8，约0.7，约0.6，约0.5，约0.4，约0.3，约0.2，约0.1。In the present application, the cation providing group includes a group that is positively charged at a pH of about 4.0 or lower. For example, the cation providing group is a group that is positively charged at the following pH conditions: about 4.0, about 3.0, about 2.0, about 1.0, about 0.9, about 0.8, about 0.7, about 0.6, about 0.5, about 0.4, about 0.3, about 0.2, about 0.1.
在本申请中，所述阳离子提供基团包含叔胺基团。其中，所述叔胺基团中与其三价基的胺连接的3个烃基各自独立地选自任选取代的取代基。In the present application, the cation providing group comprises a tertiary amine group, wherein the three hydrocarbon groups connected to the amine of the trivalent group in the tertiary amine group are independently selected from optionally substituted substituents.
在本申请中，所述氢键提供基团包含脲基。其中，所述脲基中与其2个三价基的胺分别连接的2个烃基(即总计4个烃基)各自独立地选自任选取代的取代基。In the present application, the hydrogen bond providing group comprises a urea group, wherein the two hydrocarbon groups (i.e., a total of four hydrocarbon groups) respectively connected to the amines of the two trivalent groups in the urea group are independently selected from optionally substituted substituents.
在本申请中，所述化合物包含尾部，所述尾部具有疏水性。在某些实施方式中，所述尾部取代在所述脲基的一个或两个氮原子上。在某些实施方式中，所述尾部包含饱和的脂肪链。在某些实施方式中，所述尾部包含不饱和的脂肪链。在某些实施方式中，所述尾部包含不饱和的脂肪链，且所述的不饱和为部分不饱和。在某些实施方式中，所述尾部包含不饱和的脂肪链，且所述的不饱和为单不饱和。在某些实施方式中，所述尾部包含不饱和的脂肪链，且所述的不饱和为多不饱和。In the present application, the compound comprises a tail, and the tail is hydrophobic. In certain embodiments, the tail is substituted on one or two nitrogen atoms of the urea group. In certain embodiments, the tail comprises a saturated fatty chain. In certain embodiments, the tail comprises an unsaturated fatty chain. In certain embodiments, the tail comprises an unsaturated fatty chain, and the unsaturation is partially unsaturated. In certain embodiments, the tail comprises an unsaturated fatty chain, and the unsaturation is monounsaturated. In certain embodiments, the tail comprises an unsaturated fatty chain, and the unsaturation is polyunsaturated.
在本申请中，所述尾部的主链链长为1至20个原子。例如，所述尾部具有长度为1个、2个、3个、4个、5个、6个、7个、8个、9个、10个、11个、12个、13个、14个、15个、16个、17个、18个、19个、或20个原子的主链链长。 In the present application, the main chain length of the tail is 1 to 20 atoms. For example, the tail has a main chain length of 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, or 20 atoms.
在本申请中，所述化合物包含结构(I)：其中，L1为所述氢键提供基团，所述L1任选地被取代，G1为链长为2至6个原子的连接基团，所述G1任选地被取代。例如，G1为具有以下链长的连接基团：2个、3个、4个、5个、6个原子。例如，G1选自以下任意基团或其任意组合：烃基，脂环基，脂杂环基，烷基，烯基，炔基。例如，G1任选地被取代，并且一个原子被N替换、被O替换或被S替换。In the present application, the compound comprises structure (I): Wherein, L1 is the hydrogen bond providing group, and the L1 is optionally substituted, and G1 is a linking group with a chain length of 2 to 6 atoms, and the G1 is optionally substituted. For example, G1 is a linking group with the following chain lengths: 2, 3, 4, 5, 6 atoms. For example, G1 is selected from any of the following groups or any combination thereof: hydrocarbon group, alicyclic group, alicyclic group, alkyl group, alkenyl group, alkynyl group. For example, G1 is optionally substituted, and one atom is replaced by N, replaced by O, or replaced by S.
例如，在某些实施方式中，所述化合物的结构如下(如本申请提供的结构OT13-1所述)：该化合物包含一个作为阳离子提供基团的叔胺基团，且包含所述结构(I)，其中G1为链长为3个原子的C3烷基，L1为能够作为氢键供体的脲基，并且该脲基基团的一个三价基的胺发生取代，取代基为分支化的长链烯基，其分子式为CH(C8H16CH＝CHCH2CH＝CHC5H11)2，该取代部分同时也是如本申请所定义的具有疏水性的尾部。For example, in certain embodiments, the structure of the compound is as follows (as described in structure OT13-1 provided herein): The compound comprises a tertiary amine group as a cation providing group and comprises the structure (I), wherein G1 is a C3 alkyl group with a chain length of 3 atoms, L1 is a urea group capable of serving as a hydrogen bond donor, and a trivalent amine of the urea group is substituted, the substituent is a branched long - chain alkenyl group, and the molecular formula is CH( C8H16CH = CHCH2CH = CHC5H11 ) 2 , and the substituted part is also a hydrophobic tail as defined in the present application.
在本申请中，所述化合物包含结构(II)和结构(IIIb)中的任一个：其中，G1为链长为2至6个原子的连接基团，所述G1任选地被取代，R1、R2a，R2b，R5和R6各自独立地任选地被取代。例如，G1为具有以下链长的连接基团：2个、3个、4个、5个、6个原子。例如，G1选自以下任意基团或其任意组合：烃基，脂环基，脂杂环基，烷基，烯基，炔基。例如，G1任选地被取代，并且一个原子被N替换、被O替换或被S替换。例如，R1、R2a，R2b，R5和R6各自独立地选自以下任意基团或其任意组合：烃基，酯基，酰胺，N’-烷基-N-烯基-脲基，脂环基，脂杂环基，芳基，杂芳基，C1烷基，C2烷基，C3烷基，C4烷基，C5烷基，C6烷基，C7烷基，C8烷基，C9烷基，C10烷基，C11烷基，C12烷基，C13烷基，C14烷基，C15烷基，C16烷基，C17烷基，C18烷基，C19烷基，C20烷基，C2烯基，C3烯基，C4烯基，C5烯基，C6烯基，C7烯基，C8烯基，C9烯基，C10烯基，C11烯基，C12烯基，C13烯基，C14烯基， C15烯基，C16烯基，C17烯基，C18烯基，C19烯基，C20烯基，C2炔基，C3炔基，C4炔基，C5炔基，C6炔基，C7炔基，C8炔基，C9炔基，C10炔基，C11炔基，C12炔基，C13炔基，C14炔基，C15炔基，C16炔基，C17炔基，C18炔基，C19炔基，C20炔基。例如，R1、R2a，R2b，R5和R6各自独立地为支化的或非支化的。例如，R1、R2a，R2b，R5和R6各自独立地包含0个、1个、2个、3个或4个不饱和键。例如，R1、R2a，R2b，R5和R6各自独立地被任选取代的取代基RX所取代。例如，R1、R2a，R2b，R5和R6各自独立地被任选取代的取代基RX所取代，并且RX选自以下任意基团：C1-C20烷基，C2-C25酯基，C2-C20烯基，酰胺，脲基，羟基，巯基，氨基。In the present application, the compound comprises any one of structure (II) and structure (IIIb): Wherein, G1 is a linking group with a chain length of 2 to 6 atoms, and G1 is optionally substituted, and R1 , R2a , R2b , R5 and R6 are each independently optionally substituted. For example, G1 is a linking group with the following chain lengths: 2, 3, 4, 5, 6 atoms. For example, G1 is selected from any of the following groups or any combination thereof: hydrocarbon group, alicyclic group, alicyclic group, alkyl group, alkenyl group, alkynyl group. For example, G1 is optionally substituted, and one atom is replaced by N, replaced by O or replaced by S. For example, R 1 , R 2a , R 2b , R 5 and R 6 are each independently selected from the following groups or any combination thereof: hydrocarbon group, ester group, amide group, N'-alkyl-N-alkenyl-urea group, alicyclic group, alicyclic heterocyclic group, aryl group, heteroaryl group, C 1 alkyl group, C 2 alkyl group, C 3 alkyl group, C 4 alkyl group, C 5 alkyl group, C 6 alkyl group, C 7 alkyl group, C 8 alkyl group, C 9 alkyl group, C 10 alkyl group, C 11 alkyl group, C 12 alkyl group, C 13 alkyl group, C 14 alkyl group, C 15 alkyl group, C 16 alkyl group, C 17 alkyl group, C 18 alkyl group, C 19 alkyl group, C 20 alkyl group, C 2 alkenyl group, C 3 alkenyl group, C 4 alkenyl group, C 5 alkenyl group, C 6 alkenyl group, C 7 alkenyl group, C 8 alkenyl group, C 9 alkenyl group, C 10 alkenyl group, C 11 alkenyl group, C 12 alkyl group, C 13 alkyl group, C 14 alkyl group, C 15 alkyl group, C 16 alkyl group, C 17 alkyl group, C 18 alkyl group, C 19 alkyl group, C 20 alkyl group, C 12 alkenyl, C 13 alkenyl, C 14 alkenyl, C15 alkenyl, C16 alkenyl, C17 alkenyl , C18 alkenyl, C19 alkenyl, C20 alkenyl, C2 alkynyl, C3 alkynyl, C4 alkynyl, C5 alkynyl, C6 alkynyl, C7 alkynyl, C8 alkynyl, C9 alkynyl, C10 alkynyl, C11 alkynyl, C12 alkynyl, C13 alkynyl , C14 alkynyl , C15 alkynyl , C16 alkynyl, C17 alkynyl, C18 alkynyl, C19 alkynyl , C20 alkynyl . For example, R1 , R2a , R2b , R5 and R6 are each independently branched or unbranched. For example, R 1 , R 2a , R 2b , R 5 and R 6 each independently contain 0, 1, 2, 3 or 4 unsaturated bonds. For example, R 1 , R 2a , R 2b , R 5 and R 6 are each independently substituted by an optionally substituted substituent RX . For example, R 1 , R 2a , R 2b , R 5 and R 6 are each independently substituted by an optionally substituted substituent RX , and RX is selected from any of the following groups: C 1 -C 20 alkyl, C 2 -C 25 ester, C 2 -C 20 alkenyl, amide, urea, hydroxyl, thiol, amino.
例如，在某些实施方式中，所述化合物的结构如下(如本申请提供的结构OT13-3所述)：该化合物包含1个作为阳离子提供基团的叔胺基团，且包含所述结构(II)，其中G1为链长为3个原子的C3烃基，与G1相连接的是氢键提供基团脲基，并且该脲基基团的2个三价基的胺均各自独立地发生了取代，即所述结构(II)中的R1为分支化的烷基酯类取代基，其分子式为CH(C8H16COOCH2C6H13C4H9)2，所述结构(II)中的R2a和R2b分别为氢和C8烷基(顺序不限)。For example, in certain embodiments, the structure of the compound is as follows (as described in structure OT13-3 provided herein): The compound comprises a tertiary amine group as a cation providing group and comprises the structure (II), wherein G1 is a C3 hydrocarbon group with a chain length of 3 atoms, connected to G1 is a hydrogen bond providing group urea group, and the amines of the two trivalent groups of the urea group are independently substituted, that is, R1 in the structure (II) is a branched alkyl ester substituent, and its molecular formula is CH ( C8H16COOCH2C6H13C4H9 ) 2 , and R2a and R2b in the structure (II ) are hydrogen and C8 alkyl respectively (in any order).
在本申请中，所述化合物包含结构(IIIa)和结构(IIIc)中的任一个： 其中，G1和G2各自独立地为链长为2至6个原子的连接基团，所述G1和G2各自独立地任选地被取代，R1、R2a，R2b，R3，R4a，R4b，R7，R8a，R8b和R9各自独立地任选地被取代。例如，G1和G2各自独立地为具有以下链长的连接基团：2个、3个、4个、5个、6个原子。例如，G1和G2各自独立地选自以下任意基团或其任意组合：烃基，脂环基，脂杂环基，烷基，烯基，炔基。例如，G1和G2各自独立地任选地被取代，并且一个原子被N替换、被O替换或被S替换。例如，R1、R2a，R2b，R3，R4a，R4b，R7，R8a，R8b和R9各自独立地选自以下任意基团或其任意组合：烃基，酯基，酰胺，N’-烷基-N-烯基-脲基，脂环基，脂杂环基，芳基，杂芳基，C1烷基，C2烷基，C3烷基，C4烷基，C5烷基，C6烷基，C7烷基，C8烷基，C9烷基，C10烷基，C11烷基，C12烷基，C13烷基，C14烷基，C15烷基，C16烷基，C17烷基，C18烷基，C19烷基，C20烷基，C2烯基，C3烯基，C4烯基，C5烯基，C6烯基，C7烯基，C8烯基，C9烯基，C10烯基，C11烯基，C12烯基，C13烯基，C14烯基，C15烯基，C16烯基，C17烯基，C18烯基，C19烯基，C20烯基，C2炔基，C3炔基，C4炔基，C5炔基，C6炔基，C7炔基，C8炔基，C9炔基，C10炔基，C11炔基，C12炔基，C13炔基，C14炔基，C15炔基，C16炔基，C17炔基，C18炔基，C19炔基，C20炔基。例如，R1、R2a，R2b，R3，R4a，R4b，R7，R8a，R8b和R9各自独立地为支化的或非支化的。例如，R1、R2a，R2b，R3，R4a，R4b，R7，R8a，R8b和R9各自独立地包含0个、1个、2个、3个或4个不饱和键。例如，R1、R2a，R2b，R3，R4a，R4b，R7，R8a，R8b和R9各自独立地被任选取代的取代基RX所取代。例如，R1、R2a，R2b，R3，R4a，R4b，R7，R8a，R8b和R9各自独立地被任选取代的取代基RX所取代，并且RX选自以下任意基团：C1-C20烷基，C2-C25酯基，C2-C20烯基，酰胺，脲基，羟基，巯基，氨基。In the present application, the compound comprises any one of structure (IIIa) and structure (IIIc): Wherein, G1 and G2 are each independently a linking group with a chain length of 2 to 6 atoms, and G1 and G2 are each independently optionally substituted, and R1 , R2a , R2b , R3 , R4a , R4b , R7 , R8a , R8b and R9 are each independently optionally substituted. For example, G1 and G2 are each independently a linking group with a chain length of 2, 3, 4, 5, 6 atoms. For example, G1 and G2 are each independently selected from any of the following groups or any combination thereof: hydrocarbon, alicyclic, alicyclic, alkyl, alkenyl, alkynyl. For example , G1 and G2 are each independently optionally substituted, and one atom is replaced by N, replaced by O or replaced by S. For example, R 1 , R 2a , R 2b , R 3 , R 4a , R 4b , R 7 , R 8a , R 8b and R 9 are each independently selected from the following groups or any combination thereof: a hydrocarbon group, an ester group, an amide group, an N′-alkyl-N-alkenyl-urea group, an alicyclic group, an alicyclic group, an aryl group, a heteroaryl group, a C 1 alkyl group, a C 2 alkyl group, a C 3 alkyl group, a C 4 alkyl group, a C 5 alkyl group, a C 6 alkyl group, a C 7 alkyl group, a C 8 alkyl group, a C 9 alkyl group, a C 10 alkyl group, a C 11 alkyl group, a C 12 alkyl group, a C 13 alkyl group, a C 14 alkyl group, a C 15 alkyl group, a C 16 alkyl group, a C 17 alkyl group, a C 18 alkyl group, a C 19 alkyl group, a C 20 alkyl group, a C 2 alkenyl group, a C 3 alkenyl group, a C 4 alkenyl group, a C 5 alkenyl group, a C 6 alkenyl group, a C 7 alkenyl group, a C C 8 alkenyl, C 9 alkenyl, C 10 alkenyl, C 11 alkenyl, C 12 alkenyl, C 13 alkenyl, C 14 alkenyl, C 15 alkenyl, C 16 alkenyl, C 17 alkenyl, C 18 alkenyl, C 19 alkenyl, C 20 alkenyl, C 2 alkynyl, C 3 alkynyl, C 4 alkynyl, C 5 alkynyl, C 6 alkynyl, C 7 alkynyl, C 8 alkynyl, C 9 alkynyl, C 10 alkynyl, C 11 alkynyl, C 12 alkynyl, C 13 alkynyl, C 14 alkynyl, C 15 alkynyl, C 16 alkynyl, C 17 alkynyl, C 18 alkynyl, C 19 alkynyl, C 20 alkynyl . For example, R 1 , R 2a , R 2b , R 3 , R 4a , R 4b , R 7 , R 8a , R 8b and R 9 are each independently branched or unbranched. For example, R 1 , R 2a , R 2b , R 3 , R 4a , R 4b , R 7 , R 8a , R 8b and R 9 each independently contain 0, 1, 2, 3 or 4 unsaturated bonds. For example, R 1 , R 2a , R 2b , R 3 , R 4a , R 4b , R 7 , R 8a , R 8b and R 9 are each independently substituted by an optionally substituted substituent RX . For example, R1 , R2a , R2b , R3 , R4a , R4b , R7 , R8a , R8b and R9 are each independently substituted with an optionally substituted substituent RX , and RX is selected from any of the following groups: C1 - C20 alkyl, C2 - C25 ester, C2 - C20 alkenyl, amide, urea, hydroxyl, thiol, amino.
例如，在某些实施方式中，所述化合物的结构如下(如本申请提供的结构OT13-5所述)：该化合物包含2个作为阳离子提供基团且彼此之间形成了哌嗪环(对二氮己环)的叔胺基团，且包含所述结构(IIIa)，其中 G1和G2均是链长为2个原子的C2烷基，它们分别都与氢键提供基团脲基相连接，且每个脲基各自的2个三价基的胺均各自独立地发生了取代，即所述结构(IIIa)中的R1为链长为12个原子的C12烷基，所述结构(IIIa)中的R2a和R2b分别为氢和C12烷基(顺序不限)，所述结构(IIIa)中的R3为C12烷基，所述结构(IIIa)中的R4a和R4b分别为氢和C12烷基(顺序不限)。For example, in certain embodiments, the structure of the compound is as follows (as described in structure OT13-5 provided herein): The compound comprises two tertiary amine groups as cation providing groups and forming a piperazine ring (p-diazepine ring) between each other, and comprises the structure (IIIa), wherein G1 and G2 are both C2 alkyl groups with a chain length of 2 atoms, and they are respectively connected to the hydrogen bond providing group urea group, and the amines of the two trivalent groups of each urea group are independently substituted, that is, R1 in the structure (IIIa) is a C12 alkyl group with a chain length of 12 atoms, R2a and R2b in the structure (IIIa) are hydrogen and C12 alkyl groups respectively (in any order), R3 in the structure (IIIa) is a C12 alkyl group, and R4a and R4b in the structure (IIIa) are hydrogen and C12 alkyl groups respectively (in any order).
例如，在某些实施方式中，所述化合物的结构如下(如本申请提供的结构OT13-4所述)：该化合物包含1个作为阳离子提供基团的叔胺基团，且包含所述结构(IIIc)，其中，G1和G2均是链长为6个原子的C6烷基，它们分别都与氢键提供基团脲基相连接，且每个脲基各自的直接与G1或G2的连接的三价基胺未被取代，即所述结构(IIIc)中的R1和R7均是氢，每个脲基各自的未与G1或G2直接相连接的三价基胺均各自独立地发生了取代，即所述结构(IIIc)中的R2a、R2b、R8a和R8b均是C6烷基，所述结构(IIIc)中的R9为丁醇，其分子式为C4H8OH。For example, in certain embodiments, the structure of the compound is as follows (as described in structure OT13-4 provided herein): The compound comprises a tertiary amine group as a cation providing group and comprises the structure (IIIc), wherein G1 and G2 are both C6 alkyl groups with a chain length of 6 atoms, which are respectively connected to the hydrogen bond providing group urea group, and the trivalent amine directly connected to G1 or G2 of each urea group is not substituted, that is, R1 and R7 in the structure (IIIc) are hydrogen, and the trivalent amine not directly connected to G1 or G2 of each urea group is substituted independently, that is, R2a , R2b , R8a and R8b in the structure (IIIc) are all C6 alkyl groups, and R9 in the structure (IIIc) is butanol , and its molecular formula is C4H8OH .
在本申请中，所述化合物包含结构(IIId)：其中，G1，G2，G3和G4各自 独立地为链长为2至6个原子的连接基团，所述G1，G2，G3和G4各自独立地任选地被取代，所述H1和H2各自独立地任选地被取代，所述R1、R2a，R2b，R7，R8a，R8b，R10，R11a，R11b，R12，R13a和R13b各自独立地任选地被取代。例如，G1，G2，G3和G4各自独立地为具有以下链长的连接基团：2个、3个、4个、5个、6个原子。例如，G1，G2，G3和G4各自独立地选自以下任意基团或其任意组合：烃基，脂环基，脂杂环基，烷基，烯基，炔基。例如，G1，G2，G3和G4各自独立地任选地被取代，并且一个原子被N替换、被O替换或被S替换。例如，H1和H2各自独立地选自以下任意基团或其任意组合：烃基，脂环基，脂杂环基，烷基，烯基，炔基。例如，H1和H2各自独立地任选地被取代，并且一个原子被N替换、被O替换或被S替换。例如，R1、R2a，R2b，R7，R8a，R8b，R10，R11a，R11b，R12，R13a和R13b各自独立地选自以下任意基团或其任意组合：烃基，酯基，酰胺，N’-烷基-N-烯基-脲基，脂环基，脂杂环基，芳基，杂芳基，C1烷基，C2烷基，C3烷基，C4烷基，C5烷基，C6烷基，C7烷基，C8烷基，C9烷基，C10烷基，C11烷基，C12烷基，C13烷基，C14烷基，C15烷基，C16烷基，C17烷基，C18烷基，C19烷基，C20烷基，C2烯基，C3烯基，C4烯基，C5烯基，C6烯基，C7烯基，C8烯基，C9烯基，C10烯基，C11烯基，C12烯基，C13烯基，C14烯基，C15烯基，C16烯基，C17烯基，C18烯基，C19烯基，C20烯基，C2炔基，C3炔基，C4炔基，C5炔基，C6炔基，C7炔基，C8炔基，C9炔基，C10炔基，C11炔基，C12炔基，C13炔基，C14炔基，C15炔基，C16炔基，C17炔基，C18炔基，C19炔基，C20炔基。例如，R1、R2a，R2b，R7，R8a，R8b，R10，R11a，R11b，R12，R13a和R13b各自独立地为支化的或非支化的。例如，R1、R2a，R2b，R7，R8a，R8b，R10，R11a，R11b，R12，R13a和R13b各自独立地包含0个、1个、2个、3个或4个不饱和键。例如，R1、R2a，R2b，R7，R8a，R8b，R10，R11a，R11b，R12，R13a和R13b各自独立地被任选取代的取代基RX所取代。例如，R1、R2a，R2b，R7，R8a，R8b，R10，R11a，R11b，R12，R13a和R13b各自独立地被任选取代的取代基RX所取代，并且RX选自以下任意基团：C1-C20烷基，C2-C25酯基，C2-C20烯基，酰胺，脲基，羟基，巯基，氨基。In the present application, the compound comprises structure (IIId): Among them, G1 , G2 , G3 and G4 are is independently a linking group with a chain length of 2 to 6 atoms, said G 1 , G 2 , G 3 and G 4 are each independently optionally substituted, said H 1 and H 2 are each independently optionally substituted, said R 1 , R 2a , R 2b , R 7 , R 8a , R 8b , R 10 , R 11a , R 11b , R 12 , R 13a and R 13b are each independently optionally substituted. For example, G 1 , G 2 , G 3 and G 4 are each independently a linking group with a chain length of 2, 3, 4, 5, 6 atoms. For example, G 1 , G 2 , G 3 and G 4 are each independently selected from the following any group or any combination thereof: hydrocarbon group, alicyclic group, alicyclic group, alkyl group, alkenyl group, alkynyl group. For example, G1 , G2 , G3 and G4 are each independently optionally substituted, and one atom is replaced by N, replaced by O or replaced by S. For example, H1 and H2 are each independently selected from any of the following groups or any combination thereof: hydrocarbon group, alicyclic group, alicyclic heterocyclic group, alkyl group, alkenyl group, alkynyl group. For example, H1 and H2 are each independently optionally substituted, and one atom is replaced by N, replaced by O or replaced by S. For example, R 1 , R 2a , R 2b , R 7 , R 8a , R 8b , R 10 , R 11a , R 11b , R 12 , R 13a and R 13b are each independently selected from the following groups or any combination thereof: hydrocarbon group, ester group, amide group, N'-alkyl-N-alkenyl-urea group, alicyclic group, alicyclic heterocyclic group, aryl group, heteroaryl group, C 1 alkyl group, C 2 alkyl group, C 3 alkyl group, C 4 alkyl group, C 5 alkyl group, C 6 alkyl group, C 7 alkyl group, C 8 alkyl group, C 9 alkyl group, C 10 alkyl group, C 11 alkyl group, C 12 alkyl group, C 13 alkyl group, C 14 alkyl group, C 15 alkyl group, C 16 alkyl group, C 17 alkyl group, C 18 alkyl group, C 19 alkyl group, C 20 alkyl group, C 2 alkenyl group, C 3 alkenyl group, C 4 alkenyl group, C [0014] The present invention relates to alkenyl, C5 alkenyl, C6 alkenyl, C7 alkenyl, C8 alkenyl, C9 alkenyl, C10 alkenyl, C11 alkenyl, C12 alkenyl, C13 alkenyl, C14 alkenyl, C15 alkenyl, C16 alkenyl, C17 alkenyl, C18 alkenyl, C19 alkenyl, C20 alkenyl, C2 alkynyl, C3 alkynyl, C4 alkynyl, C5 alkynyl, C6 alkynyl , C7 alkynyl , C8 alkynyl , C9 alkynyl, C10 alkynyl, C11 alkynyl , C12 alkynyl, C13 alkynyl, C14 alkynyl , C15 alkynyl, C16 alkynyl, C17 alkynyl, C18 alkynyl, C19 alkynyl, C20 alkynyl. For example, R 1 , R 2a , R 2b , R 7 , R 8a , R 8b , R 10 , R 11a , R 11b , R 12 , R 13a and R 13b are each independently branched or unbranched. For example, R 1 , R 2a , R 2b , R 7 , R 8a , R 8b , R 10 , R 11a , R 11b , R 12 , R 13a and R 13b each independently contain 0, 1, 2, 3 or 4 unsaturated bonds. For example, R 1 , R 2a , R 2b , R 7 , R 8a , R 8b , R 10 , R 11a , R 11b , R 12 , R 13a and R 13b are each independently substituted with an optionally substituted substituent RX . For example, R 1 , R 2a , R 2b , R 7 , R 8a , R 8b , R 10 , R 11a , R 11b , R 12 , R 13a and R 13b are each independently substituted with an optionally substituted substituent RX , and RX is selected from any of the following groups: C 1 -C 20 alkyl, C 2 -C 25 ester, C 2 -C 20 alkenyl, amide, urea, hydroxyl, thiol, amino.
例如，在某些实施方式中，所述化合物的结构如下(如本申请提供的结构OT13-9所述)： ，该化合物包含2个作为阳离子提供基团的叔胺基团，它们通过一段结构为3,6-双丁基哌嗪-2,5-二酮的基团相连接，该化合物还包含所述结构(IIId)，其中H1和H2均为C4烷基，G1、G2、G3和G4均是链长为2个原子的C2烷基，它们各自均与氢键提供基团脲基相连接，每个脲基各自的与G1、G2、G3或G4直接相连的三价基胺均未被取代，即所述结构(IIId)中的R1、R7、R10和R12均是氢，而每个脲基各自的未与G1、G2、G3或G4直接相连接的三价基胺均各自独立地发生了取代，即所述结构(IIId)中的R2a、R2b、R8a、R8b、R11a、R11b、R13a和R13b均是分子式为C7H14CH＝CHC7H15的长链烯基。For example, in certain embodiments, the structure of the compound is as follows (as described in structure OT13-9 provided herein): , the compound comprises two tertiary amine groups as cation providing groups, which are connected through a group with a structure of 3,6-dibutylpiperazine-2,5-dione, and the compound further comprises the structure (IIId), wherein H1 and H2 are both C4 alkyl groups, G1 , G2 , G3 and G4 are all C2 alkyl groups with a chain length of 2 atoms, and they are each connected to a hydrogen bond providing group urea group, and the trivalent amine directly connected to G1 , G2 , G3 or G4 of each urea group is not substituted, that is, R1 , R7 , R10 and R12 in the structure (IIId) are all hydrogen, and the trivalent amine not directly connected to G1 , G2 , G3 or G4 of each urea group is independently substituted, that is, R2a , R2b , R8a , R8b , R11a in the structure (IIId) , R 11b , R 13a and R 13b are all long chain alkenyl groups of the molecular formula C 7 H 14 CH＝CHC 7 H 15 .
另一方面，本申请提供了一种化合物，或其药学上可接受的盐，所述化合物的结构选自下列表格列出的结构中的任一个：



On the other hand, the present application provides a compound, or a pharmaceutically acceptable salt thereof, wherein the structure of the compound is selected from any one of the structures listed in the following table:



应理解，包含如上所示的结构(I)、(II)、或(IIIa)至(IIId)的化合物的任何实施方案，以及如上所示的化合物结构(I)、(II)、或(IIIa)至(IIId)的任何特定的取代基和/或变量可以独立地与包含结构(I)、(II)、或(IIIa)至(IIId)的化合物的其他实施方案和/或取代基和/或变量组合以形成以上未具体示出的本申请的实施方案。另外，如果在特定的实施方案和/或权利要求书中列出了任何特定的R基团、L基团、G基团、H基团或变量a、x、y或z的取代基和/或变量列表，则应理解每个单独的取代基和/或变量可以从特定的实施方案和/或权利要求书中删除，并且取代基和/或变量的其余列表将被认为在本申请的范围内。It should be understood that any embodiment of a compound comprising structures (I), (II), or (IIIa) to (IIId) as shown above, and any specific substituents and/or variables of compound structures (I), (II), or (IIIa) to (IIId) as shown above can be independently combined with other embodiments and/or substituents and/or variables of compounds comprising structures (I), (II), or (IIIa) to (IIId) to form embodiments of the present application not specifically shown above. In addition, if any specific R group, L group, G group, H group, or substituent and/or variable list of variables a, x, y, or z is listed in a specific embodiment and/or claim, it should be understood that each individual substituent and/or variable can be deleted from the specific embodiment and/or claim, and the remaining list of substituents and/or variables will be considered to be within the scope of the present application.
应理解，在本说明书中，仅当这样的贡献产生稳定的化合物时，所描绘的式的取代基和/或变量的组合才是允许的。It is understood that throughout this specification, combinations of substituents and/or variables of the depicted formulae are permissible only if such contributions result in stable compounds.
组合物combination
另一方面，本申请提供了一种组合物，所述组合物包含如本申请所述的化合物中的任一种或多种以及治疗剂。在某些实施方式下，所述组合物包含如本申请所述的化合物中的任一种或多种、治疗剂以及一种或多种选自中性脂质、类固醇和聚合物缀合的脂质的赋形剂。例如，提供一种脂质纳米颗粒(LNP)的组合物，所述LNP包含本申请所述的化合物中的任一种、治疗剂以及一种或多种选自中性脂质、类固醇和聚合物缀合的脂质的赋形剂。在某些实施方式下，所述组合物包含如本申请所述的化合物中的任一种或多种、治疗剂以及脂质体。在另一些实施方式下，所述组合物也包含其他药学上可接受的赋形剂和/或载体。On the other hand, the application provides a kind of composition, the composition comprises any one or more and therapeutic agent in the compound as described in the application.In some embodiments, the composition comprises any one or more, therapeutic agent and one or more excipients selected from the lipid of neutral lipid, steroid and polymer conjugation in the compound as described in the application.For example, a kind of composition of lipid nanoparticle (LNP) is provided, the LNP comprises any one, therapeutic agent and one or more excipients selected from the lipid of neutral lipid, steroid and polymer conjugation in the compound as described in the application.In some embodiments, the composition comprises any one or more, therapeutic agent and liposome in the compound as described in the application.In other embodiments, the composition also comprises other pharmaceutically acceptable excipients and/or carriers.
在某些实施方式下，所述组合物包含如本申请所述的化合物中的任一种或多种、治疗剂以及一种或多种选自中性脂质、类固醇和聚合物缀合的脂质的赋形剂，其中化合物与所述中性脂质的摩尔比为约2:1至约8:1。例如，提供一种包含本申请所述的化合物中的任一种或多种、治疗剂以及一种或多种选自中性脂质、类固醇和聚合物缀合的脂质的赋形剂的组合物，其中所述化合物与所述中性脂质具有如下数值范围的摩尔比：约2:1至约3:1，约2:1至约4:1，约2:1至约5:1，约2:1至约6:1，约2:1至约7:1，约2:1至约8:1，约3:1至约4:1，约3:1至约5:1，约3:1至约6:1，约3:1至约7:1，约3:1至约8:1，约4:1至约5:1，约4:1至约6:1， 约4:1至约7:1，约4:1至约8:1，约5:1至约6:1，约5:1至约7:1，约5:1至约8:1，约6:1至约7:1，约6:1至约8:1，约7:1至约8:1。In certain embodiments, the composition comprises any one or more of the compounds described herein, a therapeutic agent, and one or more excipients selected from neutral lipids, steroids, and polymer-conjugated lipids, wherein the molar ratio of the compound to the neutral lipid is about 2: 1 to about 8: 1. For example, a composition comprising any one or more of the compounds described herein, a therapeutic agent, and one or more excipients selected from neutral lipids, steroids, and polymer-conjugated lipids is provided, wherein the compound has a molar ratio of the neutral lipid in the following numerical range: about 2: 1 to about 3: 1, about 2: 1 to about 4: 1, about 2: 1 to about 5: 1, about 2: 1 to about 6: 1, about 2: 1 to about 7: 1, about 2: 1 to about 8: 1, about 3: 1 to about 4: 1, about 3: 1 to about 5: 1, about 3: 1 to about 6: 1, about 3: 1 to about 7: 1, about 3: 1 to about 8: 1, about 4: 1 to about 5: 1, about 4: 1 to about 6: 1, About 4:1 to about 7:1, about 4:1 to about 8:1, about 5:1 to about 6:1, about 5:1 to about 7:1, about 5:1 to about 8:1, about 6:1 to about 7:1, about 6:1 to about 8:1, about 7:1 to about 8:1.
在某些实施方式下，所述组合物包含如本申请所述的化合物中的任一种或多种、治疗剂以及一种多种选自中性脂质、包含胆固醇的类固醇和聚合物缀合的脂质的赋形剂。在某些实施方式下，所述组合物包含如本申请所述的化合物中的任一种或多种、治疗剂以及一种或多种选自中性脂质、类固醇和聚合物缀合的脂质的赋形剂，且所述化合物与所述类固醇具有如下数值范围的摩尔比：约5:1至约4:1，约5:1至约3:1，约5:1至约2:1，约5:1至约1:1，约4:1至约3:1，约4:1至约2:1，约4:1至约1:1，约3:1至约2:1，约3:1至约1:1，约2:1至约1:1。In certain embodiments, the composition comprises any one or more of the compounds as described herein, a therapeutic agent, and one or more excipients selected from neutral lipids, steroids containing cholesterol, and polymer-conjugated lipids. In certain embodiments, the composition comprises any one or more of the compounds as described herein, a therapeutic agent, and one or more excipients selected from neutral lipids, steroids, and polymer-conjugated lipids, and the compound and the steroid have a molar ratio in the following numerical range: about 5:1 to about 4:1, about 5:1 to about 3:1, about 5:1 to about 2:1, about 5:1 to about 1:1, about 4:1 to about 3:1, about 4:1 to about 2:1, about 4:1 to about 1:1, about 3:1 to about 2:1, about 3:1 to about 1:1, about 2:1 to about 1:1.
在某些实施方式下，所述组合物包含如本申请所述的化合物中的任一种或多种、治疗剂以及一种或多种选自中性脂质、类固醇和包含聚乙二醇化脂质的聚合物缀合的脂质的赋形剂。在某些实施方式下，所述组合物包含如本申请所述的化合物中的任一种或多种、治疗剂以及一种或多种选自中性脂质、类固醇和聚合物缀合的脂质的赋形剂，且所述化合物与所述聚合物缀合的脂质具有如下数值范围的摩尔比：约100:1至约90:1，约100:1至约80:1，约100:1至约70:1，约100:1至约60:1，约100:1至约50:1，约100:1至约40:1，约100:1至约30:1，约100:1至约20:1，约90:1至约80:1，约90:1至约70:1，约90:1至约60:1，约90:1至约50:1，约90:1至约40:1，约90:1至约30:1，约90:1至约20:1，约80:1至约70:1，约80:1至约60:1，约80:1至约50:1，约80:1至约40:1，约80:1至约30:1，约80:1至约20:1，约70:1至约60:1，约70:1至约50:1，约70:1至约40:1，约70:1至约30:1，约70:1至约20:1，约60:1至约50:1，约60:1至约40:1，约60:1至约30:1，约60:1至约20:1，约50:1至约40:1，约50:1至约30:1，约50:1至约20:1，约40:1至约30:1，约40:1至约20:1，约30:1至约20:1。In certain embodiments, the composition comprises any one or more of the compounds described herein, a therapeutic agent, and one or more excipients selected from a neutral lipid, a steroid, and a polymer-conjugated lipid including a pegylated lipid. In certain embodiments, the composition comprises any one or more of the compounds as described herein, a therapeutic agent, and one or more excipients selected from a neutral lipid, a steroid, and a polymer-conjugated lipid, and the compound has a molar ratio to the polymer-conjugated lipid in the following numerical range: from about 100:1 to about 90:1, from about 100:1 to about 80:1, from about 100:1 to about 70:1, from about 100:1 to about 60:1, from about 100:1 to about 50:1, from about 100:1 to about 40:1, from about 100:1 to about 30:1, from about 100:1 to about 20:1, from about 90:1 to about 80:1, from about 90:1 to about 70:1, from about 90:1 to about 60:1, from about 90:1 to about 50:1, from about 90:1 to about 40:1, from about 90:1 to about about 30:1, about 90:1 to about 20:1, about 80:1 to about 70:1, about 80:1 to about 60:1, about 80:1 to about 50:1, about 80:1 to about 40:1, about 80:1 to about 30:1, about 80:1 to about 20:1, about 70:1 to about 60:1, about 70:1 to about 50:1, about 70:1 to about 40:1, about 70:1 to about 30:1, about 70:1 to about 20:1, about 60:1 to about 50:1, about 60:1 to about 40:1, about 60:1 to about 30:1, about 60:1 to about 20:1, about 50:1 to about 40:1, about 50:1 to about 30:1, about 50:1 to about 20:1, about 40:1 to about 30:1, about 40:1 to about 20:1, about 30:1 to about 20:1.
在本申请中，所述组合物的直径约为500nm或更小。例如，所述组合物具有如下列数值的直径大小：约500nm，约450nm，约400nm，约350nm，约300nm，约250nm，约200nm，约150nm，约100nm，约75nm，约50nm，约25nm。In the present application, the diameter of the composition is about 500nm or less. For example, the composition has a diameter of the following values: about 500nm, about 450nm, about 400nm, about 350nm, about 300nm, about 250nm, about 200nm, about 150nm, about 100nm, about 75nm, about 50nm, about 25nm.
在本申请中，所述治疗剂包含一种或多种核酸。应理解，本申请公开的内容不限于本说明书所公开的具体核酸，并且在范围上不限于核酸的任何特定来源、序列或类型，但本领域普通技术人员可以轻易地鉴定各种其它核酸来源中的相关同系物，包括来自非人类物种的核酸。预期在本申请公开的内容中使用的核酸可以包含基于天然存在的序列的序列。在某些实施方式下，所述治疗剂包含一种或多种核酸，所述核酸包含一个或多个经修饰的核苷(例如，包含一个或多个经糖部分修饰的核苷)，以期包含该核酸的化合物可能具有期望的特性。在某 些实施方式下，所述治疗剂包含一种或多种核酸，所述核酸为包含经修饰的核苷(例如，包含经修饰的糖、经修饰的核苷碱基、经修饰的键和/或经修饰的其他位置如3’末端核苷酸上的糖的3’位置和/或5’末端核苷酸的5’位置的核苷酸)的寡核苷酸，通过选择某种特定修饰以期得到的寡核苷酸可能具有期望的特征。In the present application, the therapeutic agent comprises one or more nucleic acids. It should be understood that the content disclosed in this application is not limited to the specific nucleic acids disclosed in this specification, and is not limited in scope to any specific source, sequence or type of nucleic acid, but a person of ordinary skill in the art can easily identify related homologs in various other nucleic acid sources, including nucleic acids from non-human species. It is contemplated that the nucleic acids used in the content disclosed in this application may comprise sequences based on naturally occurring sequences. In certain embodiments, the therapeutic agent comprises one or more nucleic acids, and the nucleic acid comprises one or more modified nucleosides (for example, comprising one or more nucleosides modified with a sugar moiety), in the hope that the compound comprising the nucleic acid may have desired properties. In a certain In some embodiments, the therapeutic agent comprises one or more nucleic acids that are oligonucleotides comprising modified nucleosides (e.g., oligonucleotides comprising modified sugars, modified nucleobases, modified bonds, and/or modified nucleotides at other positions such as the 3' position of the sugar on the 3' terminal nucleotide and/or the 5' position of the 5' terminal nucleotide), and the oligonucleotides obtained by selecting a particular modification may have desired characteristics.
在本申请中，所述治疗剂包含CRISPR系统的组分和/或编码Cas酶的核酸。所述“CRISPR系统”也可称为“CRISPR/Cas系统”，是一种用于生物体中位点特异性基因组靶向的工具。例如，它可以是II型CRISPR/Cas系统，它是一种原核适应性免疫反应系统，该系统使用非编码RNA指导Cas核酸酶诱导位点特异性DNA切割，并随之引发细胞DNA修复机制，即通过非同源末端连接DNA修复途径(NHEJ)或同源性定向修复(HDR)途径修复DNA损伤；该系统也可以被递送至哺乳动物细胞或真核细胞中介导基因组编辑(如，基因敲除或敲入)。例如，所述治疗剂可以包含一种或多种Cas核酸酶，其非限制性例子包括Cas1、Cas1B、Cas2、Cas3、Cas4、Cas5、Cas6、Cas7、Cas8、Cas9(也被称作Csn1和Csx12)、Cas10、Csy1、Csy2、Csy3、Cse1、Cse2、Csc1、Csc2、Csa5、Csn2、Csm2、Csm3、Csm4、Csm5、Csm6、Cmr1、Cmr3、Cmr4、Cmr5、Cmr6、Csb1、Csb2、Csb3、Csx17、Csx14、Csx10、Csx16、CsaX、Csx3、Csx1、Csx15、Csf1、Csf2、Csf3、Csf4、其同系物或其修饰形式，这些酶是已知的，本领域技术人员可以轻易从例如SwissProt数据库中找到。例如，所述治疗剂可以包含Cas9酶和/或其突变形式，以用于单链或双链的靶多核苷酸在靶序列位置的特异性结合和/或定点切割。例如，所述治疗剂包含编码CRISPR系统的组分的核酸，所述“编码”是指目标多核苷酸序列能够根据在宿主细胞/宿主动物中起作用的遗传密码被转录和/或翻译以产生相应的基因产物(例如核酸酶)。例如，所述治疗剂包含指导RNA(guide RNA或gRNA)。例如，所述治疗剂包含编码Cas酶的核酸。在某些实施方式下，所述“编码Cas酶的核酸”包含DNA(例如质粒)或RNA(例如mRNA)。In the present application, the therapeutic agent comprises components of the CRISPR system and/or nucleic acids encoding Cas enzymes. The "CRISPR system" may also be referred to as a "CRISPR/Cas system", which is a tool for site-specific genome targeting in an organism. For example, it may be a type II CRISPR/Cas system, which is a prokaryotic adaptive immune response system that uses non-coding RNA to guide Cas nucleases to induce site-specific DNA cleavage, and subsequently triggers cellular DNA repair mechanisms, i.e., repairing DNA damage through non-homologous end-joining DNA repair pathways (NHEJ) or homology-directed repair (HDR) pathways; the system may also be delivered to mammalian cells or eukaryotic cells to mediate genome editing (e.g., gene knockout or knock-in). For example, the therapeutic agent can include one or more Cas nucleases, non-limiting examples of which include Cas1, Cas1B, Cas2, Cas3, Cas4, Cas5, Cas6, Cas7, Cas8, Cas9 (also known as Csn1 and Csx12), Cas10, Csy1, Csy2, Csy3, Cse1, Cse2, Csc1, Csc2, Csa5, Csn2, Csm2, Csm3, Csm4 , Csm5, Csm6, Cmr1, Cmr3, Cmr4, Cmr5, Cmr6, Csb1, Csb2, Csb3, Csx17, Csx14, Csx10, Csx16, CsaX, Csx3, Csx1, Csx15, Csf1, Csf2, Csf3, Csf4, homologs thereof or modified forms thereof, these enzymes are known and can be easily found by a person skilled in the art from, for example, the SwissProt database. For example, the therapeutic agent may comprise a Cas9 enzyme and/or a mutant form thereof for specific binding and/or site-directed cleavage of a single-stranded or double-stranded target polynucleotide at a target sequence position. For example, the therapeutic agent comprises a nucleic acid encoding a component of a CRISPR system, wherein “encoding” means that the target polynucleotide sequence can be transcribed and/or translated according to a genetic code that functions in a host cell/host animal to produce a corresponding gene product (e.g., a nuclease). For example, the therapeutic agent comprises a guide RNA (guide RNA or gRNA). For example, the therapeutic agent comprises a nucleic acid encoding a Cas enzyme. In certain embodiments, the "nucleic acid encoding a Cas enzyme" comprises DNA (e.g., a plasmid) or RNA (e.g., mRNA).
细胞及试剂盒Cells and kits
另一方面，本申请提供了一种细胞，所述细胞包含如本申请所述的化合物和/或组合物。例如，所述细胞为一种载体细胞，其包含带负电荷的细胞膜。例如，本申请所述的包含有所述化合物的LNP在合适的pH环境下发生质子化，进而与所述载体细胞的细胞膜通过静电相互作用而聚集在一起。例如，本申请所述的化合物和/或组合物在合适的pH环境下发生质子化，使得所述载体细胞的细胞膜介导内吞作用或膜渗透作用而将所述化合物和/或组合物包含在内。On the other hand, the present application provides a cell comprising a compound and/or composition as described in the present application. For example, the cell is a carrier cell comprising a negatively charged cell membrane. For example, the LNP comprising the compound described in the present application is protonated under a suitable pH environment, and then aggregated with the cell membrane of the carrier cell through electrostatic interaction. For example, the compound and/or composition described in the present application is protonated under a suitable pH environment, so that the cell membrane of the carrier cell mediates endocytosis or membrane permeation to include the compound and/or composition.
另一方面，本申请提供了一种试剂盒，所述试剂盒包含本申请所述的化合物、本申请所 述的组合物和/或本申请所述的细胞。例如，所述试剂盒包括至少一个小瓶、管、烧瓶、瓶子、注射器或其它合适的容器，其内可以放置组分。例如，在所述试剂盒中存在超过一种组分的情况下，试剂盒还将包含有第二个、第三个或其它额外的容器，其内可以分开放置额外的组分。例如，可以在容器中包含组分的各种组合。例如，所有LNP组分都组合在单个容器中。例如，一些或所有的LNP组分提供在分开的容器中。在某些实施方式下，所述试剂盒还将包括用于容纳各种容器的包装。例如，所述试剂盒包括关于使用试剂盒组分的说明书。例如，所述说明书包括纸质的实体形式和/或可机读的电子形式。On the other hand, the present application provides a kit, the kit comprising the compound described in the present application, The composition and/or the cell described in the present application. For example, the kit includes at least one vial, tube, flask, bottle, syringe or other suitable container, in which the components can be placed. For example, when there is more than one component in the kit, the kit will also include a second, third or other additional container, in which the additional components can be placed separately. For example, various combinations of components can be included in the container. For example, all LNP components are combined in a single container. For example, some or all of the LNP components are provided in separate containers. In certain embodiments, the kit will also include packaging for accommodating various containers. For example, the kit includes instructions for using the kit components. For example, the instructions include a paper-based physical form and/or a machine-readable electronic form.
施用治疗剂的方法Methods of administering therapeutic agents
另一方面，本申请提供了一种施用治疗剂的方法，所述方法包含提供本申请所述的化合物、本申请所述的组合物、本申请所述的细胞和/或本申请所述的试剂盒。在某些实施方式下，所述方法包括将本申请所述的治疗剂与本申请所述的化合物混合。在另一些实施方式下，所述方法包括将本申请所述的治疗剂置于本申请所述的组合物中、本申请所述的细胞中和/或本申请所述的试剂盒中。On the other hand, the present application provides a method of administering a therapeutic agent, the method comprising providing a compound described herein, a composition described herein, a cell described herein, and/or a kit described herein. In certain embodiments, the method comprises mixing the therapeutic agent described herein with the compound described herein. In other embodiments, the method comprises placing the therapeutic agent described herein in a composition described herein, a cell described herein, and/or a kit described herein.
递送核酸的方法Methods of delivering nucleic acids
另一方面，本申请提供了一种递送核酸的方法，所述方法包含提供本申请所述的化合物、本申请所述的组合物、本申请所述的细胞和/或本申请所述的试剂盒。在某些实施方式下，所述方法包括将本申请所述的核酸与本申请所述的化合物混合。在另一些实施方式下，所述方法包括将本申请所述的核酸置于本申请所述的组合物中、本申请所述的细胞中和/或本申请所述的试剂盒中。例如，所述递送核酸的方法包含对本申请所述的化合物、本申请所述的组合物、本申请所述的细胞和/或本申请所述的试剂盒进行全身递送，使其广泛暴露于身体的大部分，可以通过本领域已知的任何手段进行，包括但不限于静脉内、动脉内、皮下和腹膜内递送。例如，所述递送核酸的方法包含对本申请所述的化合物、本申请所述的组合物、本申请所述的细胞和/或本申请所述的试剂盒进行局部递送，使其直接到达有机体内的靶部位，可以通过例如直接注射到疾病部位(如，肿瘤或炎症部位)或靶器官(如，肝脏、心脏、胰腺、肾脏等)中来进行。例如，所述局部递送包括局部施用或局部注射技术，包括但不限于肌内、皮下或皮内注射。例如，所述局部递送不排除全身性的药理学作用。On the other hand, the present application provides a method for delivering nucleic acid, the method comprising providing a compound described in the present application, a composition described in the present application, a cell described in the present application, and/or a kit described in the present application. In some embodiments, the method comprises mixing the nucleic acid described in the present application with the compound described in the present application. In other embodiments, the method comprises placing the nucleic acid described in the present application in a composition described in the present application, a cell described in the present application, and/or a kit described in the present application. For example, the method for delivering nucleic acid comprises delivering the compound described in the present application, the composition described in the present application, the cell described in the present application, and/or the kit described in the present application to the whole body, so that it is widely exposed to most of the body, and can be carried out by any means known in the art, including but not limited to intravenous, intra-arterial, subcutaneous and intraperitoneal delivery. For example, the method for delivering nucleic acid comprises delivering the compound described in the present application, the composition described in the present application, the cell described in the present application, and/or the kit described in the present application to the local area, so that it directly reaches the target site in the organism, and can be carried out by, for example, direct injection into the disease site (such as, tumor or inflammation site) or target organ (such as, liver, heart, pancreas, kidney, etc.). For example, the local delivery includes local administration or local injection technology, including but not limited to intramuscular, subcutaneous or intradermal injection. For example, such local delivery does not preclude systemic pharmacological effects.
治疗和/或预防疾病或病症的方法Methods of treating and/or preventing diseases or conditions
另一方面，本申请提供了一种药物组合物，其包含本申请所述的化合物、本申请所述的组合物、本申请所述的细胞和/或本申请所述的试剂盒。在某些实施方式下，所述药物组合物还包含药学上可接受的辅助成分。在某些实施方式下，所述药物组合物用于治疗和/或预防疾 病。On the other hand, the present application provides a pharmaceutical composition, which comprises the compound described in the present application, the composition described in the present application, the cell described in the present application and/or the kit described in the present application. In certain embodiments, the pharmaceutical composition further comprises a pharmaceutically acceptable auxiliary component. In certain embodiments, the pharmaceutical composition is used to treat and/or prevent a disease. sick.
另一方面，本申请提供了本申请所述的化合物、本申请所述的组合物、本申请所述的细胞、本申请所述的试剂盒和/或本申请所述的药物组合物用于制备药物的用途，所述药物用于治疗或预防疾病。On the other hand, the present application provides the use of the compounds described herein, the compositions described herein, the cells described herein, the kits described herein and/or the pharmaceutical compositions described herein for preparing drugs for treating or preventing diseases.
另一方面，本申请提供了本申请所述的化合物、本申请所述的组合物、本申请所述的细胞、本申请所述的试剂盒和/或本申请所述的药物组合物，其用于治疗或预防疾病。On the other hand, the present application provides the compounds described herein, the compositions described herein, the cells described herein, the kits described herein and/or the pharmaceutical compositions described herein, which are used for treating or preventing diseases.
另一方面，本申请提供了一种治疗和/或预防疾病的方法，其包括向受试者或患者施用合适剂量的本申请所述的化合物、本申请所述的组合物、本申请所述的细胞和/或本申请所述的试剂盒。On the other hand, the present application provides a method for treating and/or preventing a disease, which comprises administering to a subject or patient a suitable dose of the compound described herein, the composition described herein, the cell described herein and/or the kit described herein.
在本申请中，所述疾病包含癌症。In the present application, the disease comprises cancer.
在本申请中，所述疾病包含与蛋白质或酶缺乏有关的疾病。In the present application, the diseases include diseases associated with protein or enzyme deficiency.
在本申请中，所述疾病包含与特定miRNA或一组以及一组以上miRNA相关的疾病。In the present application, the disease includes diseases associated with a specific miRNA or a group or more than one group of miRNAs.
不欲被任何理论所限，下文中的实施例仅仅是为了阐释本申请的化合物、制备方法和用途等，而不用于限制本申请发明的范围。Without intending to be bound by any theory, the following examples are merely intended to illustrate the compounds, preparation methods and uses of the present application, and are not intended to limit the scope of the present invention.
实施例Example
实施例1化合物的制备Example 1 Preparation of Compounds
OT13-1的合成步骤Synthesis steps of OT13-1
1.(6E,10E)-18-溴代十八烷-6,9-二烯((6E,9E)-18-bromooctadeca-6,9-diene)的合成
1. Synthesis of (6E,10E)-18-bromooctadeca-6,9-diene
室温下将十八烷-9,2-二烯-1-基甲烷磺酸(octadeca-9,12-dien-1-yl methanesulfonate)和溴化镁溶解于THF中，并加入乙醚，随后搅拌12小时。旋蒸去除有机溶剂，水洗产物。Octadeca-9,12-dien-1-yl methanesulfonate and magnesium bromide were dissolved in THF at room temperature, and ether was added, followed by stirring for 12 hours. The organic solvent was removed by rotary evaporation, and the product was washed with water.
2.(6E,10E,28E,31E)-三十七-6,9,28,31-四烯-19-醇((6E,9E,28E,31E)-heptatriaconta-6,9,28,31-tetraen-19-ol)的合成
2. Synthesis of (6E,10E,28E,31E)-heptatriaconta-6,9,28,31-tetraen-19-ol
将(6E,9E)-18-溴代十八烷-6,9-二烯(步骤1.的产物)溶解于含有镁、碘的THF作为反应溶剂，加入甲酸乙酯，搅拌反应12小时Dissolve (6E,9E)-18-bromooctadecane-6,9-diene (the product of step 1.) in THF containing magnesium and iodine as the reaction solvent, add ethyl formate, and stir the reaction for 12 hours.
3.(6E,10E,28E,31E)-三十七-6,9,28,31-四烯-19-胺((6E,9E,28E,31E)-heptatriaconta-6,9,28,31-tetraen-19-amine)的合成
3. Synthesis of (6E,10E,28E,31E)-heptatriaconta-6,9,28,31-tetraen-19-amine
将(6E,10E,28E,31E)-三十七-6,9,28,31-四烯-19-醇(步骤2.的产物)和PCC(吡啶氯铬酸盐)在二氯甲烷里0-20℃反应两个小时，之后室温下加入氰基硼氢化钠，持续搅拌超过2小时。产物经矽鈣石过滤，并且用热乙酸乙酯洗涤。(6E,10E,28E,31E)-heptatriacontahedral-6,9,28,31-tetraen-19-ol (the product of step 2.) and PCC (pyridinium chlorochromate) were reacted in dichloromethane at 0-20°C for two hours, and then sodium cyanoborohydride was added at room temperature and stirred for more than 2 hours. The product was filtered through silica and washed with hot ethyl acetate.
4.OT13-1的合成
4. Synthesis of OT13-1
室温下将(6E,10E,28E,31E)-三十七-6,9,28,31-四烯-19-胺(步骤3.的产物)溶解于THF。加入CDI(N,N'-羰基二咪唑)并加热到70℃，通入新戊烷，持续搅拌两小时。产物用水洗后用硅胶色谱柱纯化。Dissolve (6E,10E,28E,31E)-triacontahedrine-6,9,28,31-tetraen-19-amine (the product of step 3) in THF at room temperature. Add CDI (N,N'-carbonyldiimidazole) and heat to 70°C, pass neopentane, and continue stirring for two hours. Wash the product with water and purify it with a silica gel column.
OT13-2的合成步骤Synthesis steps of OT13-2
5.甲烷磺酸-(10E,12E)-十八-9,12-二烯-1-基酯((9E,12E)-octadeca-9,12-dien-1-yl methanesulfonate)的合成
5. Synthesis of (10E,12E)-octadeca-9,12-dien-1-yl methanesulfonate
室温下，将MsCl(甲磺酰氯)溶解于二氯甲烷中，在惰性气体条件下，将(10E,12E)-十八-9,12-二烯-1-醇((9E,12E)-octadeca-9,12-dien-1-ol)与甲磺酰氯在0-20℃反应温度及三乙胺催化下进行温和搅拌，生成铵盐。用蒸馏水终止反应，并洗涤总有机层，再用饱和卤盐洗涤，干燥，浓缩。用硅胶柱色谱纯化直至纯度大于85％。At room temperature, MsCl (methanesulfonyl chloride) was dissolved in dichloromethane. Under inert gas conditions, (10E,12E)-octadeca-9,12-dien-1-ol ((9E,12E)-octadeca-9,12-dien-1-ol) was gently stirred with methanesulfonyl chloride at a reaction temperature of 0-20°C and catalyzed by triethylamine to generate ammonium salt. The reaction was terminated with distilled water, and the total organic layer was washed with saturated halide, dried, and concentrated. Purification was performed by silica gel column chromatography until the purity was greater than 85%.
6. 2-[(10E,12E)-十八-9,12-二烯基]异吲哚-1,3-二酮(2-((9E,12E)-octadeca-9,12-dien-1-yl)isoindoline-1,3-dione)的合成
6. Synthesis of 2-[(10E,12E)-octadeca-9,12-dien-1-yl]isoindoline-1,3-dione
将甲烷磺酸-(10E,12E)-十八-9,12-二烯-1-基酯(步骤5.的产物)，邻苯二甲酰亚胺钾盐溶解于二甲基甲酰胺，并在室温下加入叠氮化钠。在70℃反应温度下持续搅拌2小时，减压蒸馏。加入二氯甲烷萃取，并用水洗涤，经无水硫酸钠干燥后得到粗产物，并用色谱柱进行分 离纯化。Dissolve methanesulfonic acid (10E, 12E)-octadecane-9,12-diene-1-yl ester (the product of step 5) and potassium phthalimide in dimethylformamide, and add sodium azide at room temperature. Continue stirring at 70°C for 2 hours and distill under reduced pressure. Add dichloromethane to extract, wash with water, and dry over anhydrous sodium sulfate to obtain a crude product, which is then separated by chromatography. Purification.
7.(10E,12E)-十八-9,12-二烯-1-胺((9E,12E)-octadeca-9,12-dien-1-amine)的合成
7. Synthesis of (10E,12E)-octadeca-9,12-dien-1-amine
将含水肼加入乙醇并与2-[(10E,12E)-十八-9,12-二烯基]异吲哚-1,3-二酮(步骤6.的产物)在惰性气体，0-20℃下反应过两个小时。将产物干燥并用硅胶色谱柱纯化。Aqueous hydrazine was added to ethanol and reacted with 2-[(10E,12E)-octadeca-9,12-dienyl]isoindole-1,3-dione (the product of step 6) under inert gas at 0-20° C. for two hours. The product was dried and purified by silica gel column chromatography.
8.双[(6E,10E)-十八-6,9-二烯-18-基]胺(di((9E,12E)-octadeca-9,12-dien-1-yl)amine)的合成
8. Synthesis of di((9E,12E)-octadeca-9,12-dien-1-yl)amine
在碱性条件下(pH在8-8.5之间)，缓慢加入甲烷磺酸-(10E,12E)-十八-9,12-二烯-1-基酯(步骤5.的产物)，直至溶解。加热至100℃并持续搅拌2小时。通过色谱监测反应，将反应物冷却至室温加入水，用乙酸乙酯萃取水层化合物。将产物用水洗涤再用PBS盐溶液洗涤，产物经无水Na2SO4干燥并浓缩。Under alkaline conditions (pH between 8-8.5), slowly add methanesulfonic acid-(10E,12E)-octadec-9,12-diene-1-yl ester (the product of step 5.) until dissolved. Heat to 100°C and continue stirring for 2 hours. Monitor the reaction by chromatography, cool the reactant to room temperature, add water, and extract the aqueous layer compound with ethyl acetate. Wash the product with water and then with PBS saline solution, dry the product over anhydrous Na 2 SO 4 and concentrate.
9.OT13-2的合成
9. Synthesis of OT13-2
室温下将双[(6E,10E)-十八-6,9-二烯-18-基]胺(步骤8.的产物)溶解于THF。加入CDI(N,N'-羰基二咪唑)并加热到70℃，通入新戊烷，持续搅拌两小时。产物用水洗后用硅胶色谱柱纯化。Dissolve bis[(6E,10E)-octadecane-6,9-dien-18-yl]amine (the product of step 8) in THF at room temperature. Add CDI (N,N'-carbonyldiimidazole) and heat to 70°C, pass neopentane, and continue stirring for two hours. Wash the product with water and purify it with a silica gel column.
OT13-3的合成步骤Synthesis steps of OT13-3
10. 19-乙氧基-9,11-双(乙氧基羰基)-10,19-二氧亚基十九烷酸乙酯(tetraethyl 9-oxoheptadecane-1.8,10,17-tetracarboxylate)的合成
10. Synthesis of 19-ethoxy-9,11-bis(ethoxycarbonyl)-10,19-dioxynonadecanoic acid ethyl ester (tetraethyl 9-oxoheptadecane-1.8,10,17-tetracarboxylate)
白色粉末1,3-丙酮二羧酸二乙酯加入含8-溴辛酸乙酯和乙氧化钠的纯乙醇中，在80℃下反应48小时。冰浴条件下用甲醇洗涤，产物用硅胶色谱柱纯化。White powder 1,3-acetone dicarboxylic acid diethyl ester was added to pure ethanol containing 8-bromooctanoic acid ethyl ester and sodium ethoxide, and reacted at 80°C for 48 hours. The product was washed with methanol under ice bath conditions and purified by silica gel column chromatography.
11. 10-氧亚基十九烷二酸(10-oxononadecanedioic acid)的合成
11. Synthesis of 10-oxononadecanedioic acid
19-乙氧基-9,11-双(乙氧基羰基)-10,19-二氧亚基十九烷酸乙酯(步骤10.的产物)与12M浓盐酸和乙酸在110℃下反应超过12小时，得到产物经过饱和氯化钠溶液洗涤，用无水硫酸钠干燥，减压蒸馏去除剩余溶剂，得到产物再经过硅胶色谱柱纯化。19-Ethoxy-9,11-bis(ethoxycarbonyl)-10,19-dioxyylidene nonadecanoic acid ethyl ester (the product of step 10.) was reacted with 12M concentrated hydrochloric acid and acetic acid at 110°C for more than 12 hours to obtain a product which was washed with a saturated sodium chloride solution, dried over anhydrous sodium sulfate, and the remaining solvent was removed by distillation under reduced pressure to obtain a product which was then purified by silica gel chromatography.
12. 19-[(2-丁基辛基)氧基]-10,19-二氧亚基十九烷酸-2-丁基辛基酯(bis(2-butyloctyl)10-oxononadecanedioate)的合成
12. Synthesis of bis(2-butyloctyl)10-oxononadecanedioate
10-氧亚基十九烷二酸(步骤11.的产物)和2-丁基辛醇在EDCI,DMAP和DCM的催化下在30℃下进行12小时酯化反应。分离有机层，用饱和氯化钠溶液洗涤，减压去除剩余溶剂，得到产物用硅胶色谱柱进行纯化。10-Oxylidene nonadecanedioic acid (product of step 11.) and 2-butyloctanol were subjected to esterification reaction at 30°C for 12 hours under the catalysis of EDCI, DMAP and DCM. The organic layer was separated and washed with saturated sodium chloride solution, and the remaining solvent was removed under reduced pressure to obtain the product, which was purified by silica gel chromatography.
13. 19-[(2-丁基辛基)氧基]-10-{[3-(二甲基氨基)丙基]氨基}-19-氧亚基十九烷酸-2-丁基辛基酯(bis(2-butyloctyl)10-((3-(dimethylamino)propyl)amino)nonadecanedioate)的合成
13. Synthesis of bis(2-butyloctyl)10-((3-(dimethylamino)propyl)amino)nonadecanedioate
19-[(2-丁基辛基)氧基]-10,19-二氧亚基十九烷酸-2-丁基辛基酯(步骤12.的产物)和3-(二甲基氨基)丙-1-胺(N-dimethylpropane-1,3-diamine)加入含有乙酸和三乙酰氧基硼氢化钠的二氯乙烯里，在40℃下搅拌反应12小时。冷却到室温后，加入水和乙酸乙酯，将产物萃取到乙酸乙酯中。有机层减压蒸发，真空干燥浓缩。19-[(2-butyloctyl)oxy]-10,19-dioxyylidene nonadecanoic acid-2-butyloctyl ester (product of step 12.) and 3-(dimethylamino)propane-1-amine (N-dimethylpropane-1,3-diamine) were added to ethylene dichloride containing acetic acid and sodium triacetoxyborohydride, and stirred at 40°C for 12 hours. After cooling to room temperature, water and ethyl acetate were added, and the product was extracted into ethyl acetate. The organic layer was evaporated under reduced pressure, dried and concentrated in vacuo.
14.OT13-3的合成
14. Synthesis of OT13-3
19-[(2-丁基辛基)氧基]-10-{[3-(二甲基氨基)丙基]氨基}-19-氧亚基十九烷酸-2-丁基辛基酯 (步骤13.的产物)室温下在CDI的催化下同1-辛胺在THF中通入新戊烷，搅拌反应2小时。产物用水洗后用硅胶色谱柱纯化。2-Butyloctyl 19-[(2-butyloctyl)oxy]-10-{[3-(dimethylamino)propyl]amino}-19-oxyylidene nonadecanoate (Product of step 13.) Under the catalysis of CDI, 1-octylamine was added to THF and neopentane was introduced at room temperature, and the reaction was stirred for 2 hours. The product was washed with water and purified by silica gel column chromatography.
OT13-4的合成步骤Synthesis steps of OT13-4
15. 6-溴己-1-胺(6-bromohexan-1-amine)的合成
15. Synthesis of 6-bromohexan-1-amine
圆底烧瓶中热氢溴酸溶解6-氨基-1-己醇，不高于70℃沸腾反应两小时。用乙酸乙酯萃取，杂质经碳酸钠去除，产物之后水洗并干燥。Dissolve 6-amino-1-hexanol in hot hydrobromic acid in a round-bottom flask and boil at no higher than 70°C for two hours. Extract with ethyl acetate, remove impurities with sodium carbonate, and then wash with water and dry.
16. 1-[(6-溴己基)氨基]-N,N-二己基甲烷酰胺(3-(6-bromohexyl)-1,1-dihexylurea)的合成
16. Synthesis of 1-[(6-bromohexyl)amino]-N,N-dihexylmethaneamide (3-(6-bromohexyl)-1,1-dihexylurea)
6-溴己-1-胺(步骤15.的产物)在CDI的催化下与二己胺在THF下加热至70℃，通入新戊烷，震荡反应2小时。产物由硅胶色谱柱纯化。6-Bromohexan-1-amine (the product of step 15) was heated to 70°C in THF with dihexylamine under the catalysis of CDI, and neopentane was introduced and the reaction was shaken for 2 hours. The product was purified by silica gel column chromatography.
17.OT13-4的合成
17. Synthesis of OT13-4
在25-70℃含碳酸钠无水乙醇条件，1-[(6-溴己基)氨基]-N,N-二己基甲烷酰胺(步骤16.的产物)和4-氨基-1-丁醇振荡反应2小时。旋蒸除有机溶剂，经水洗干燥并用硅胶色谱柱纯化。室温下将双[(6E,10E)-十八-6,9-二烯-18-基]胺(步骤8.的产物)溶解于THF。加入CDI(N,N'-羰基二咪唑)并加热到70℃，通入新戊烷，持续搅拌两小时。产物用水洗后用硅胶色谱柱纯化。1-[(6-bromohexyl)amino]-N,N-dihexylmethaneamide (the product of step 16.) and 4-amino-1-butanol were shaken and reacted for 2 hours in anhydrous ethanol containing sodium carbonate at 25-70°C. The organic solvent was removed by rotary evaporation, and the mixture was washed with water, dried and purified by silica gel chromatography. Di[(6E,10E)-octadecane-6,9-diene-18-yl]amine (the product of step 8.) was dissolved in THF at room temperature. CDI (N,N'-carbonyldiimidazole) was added and heated to 70°C, and neopentane was introduced and stirred for two hours. The product was washed with water and purified by silica gel chromatography.
OT13-5、OT13-6和OT13-7的合成步骤Synthesis steps of OT13-5, OT13-6 and OT13-7
18. 1-(十二烷基氨基)-N-{2-[4-(4-氧亚基-3,5-二氮杂十七烷-1-基)哌嗪-1-基]乙基}甲烷酰胺(1,1’-(piperazine-1,4-diylbis(ethane-2,1-diyl)bis(3-dodecylurea))的合成
18. Synthesis of 1-(dodecylamino)-N-{2-[4-(4-oxyylidene-3,5-diazaheptadecan-1-yl)piperazin-1-yl]ethyl}methaneamide (1,1'-(piperazine-1,4-diylbis(ethane-2,1-diyl)bis(3-dodecylurea))
2-[4-(2-氨基乙基)哌嗪-1-基]乙-1-胺在CDI的催化下与十二烷-1-胺在THF下加热至70℃，通入新戊烷，震荡反应2小时。产物由硅胶色谱柱纯化，得到浅黄色固体。2-[4-(2-aminoethyl)piperazin-1-yl]ethan-1-amine was catalyzed by CDI and heated to 70°C in THF, and then introduced with neopentane and shaken for 2 hours. The product was purified by silica gel column chromatography to obtain a light yellow solid.
19.OT13-5的合成
19. Synthesis of OT13-5
1-(十二烷基氨基)-N-{2-[4-(4-氧亚基-3,5-二氮杂十七烷-1-基)哌嗪-1-基]乙基}甲烷酰胺(步骤18.的产物)和环氧12烷混合，在含有氯乙腈的THF中密封80℃加热持续搅拌反应2天，直至MALDI-TOF质谱检测到目标产物。粗产物经由硅胶色谱纯化，产物最终由核磁确认。1-(Dodecylamino)-N-{2-[4-(4-oxyylidene-3,5-diazaheptadecan-1-yl)piperazin-1-yl]ethyl}methaneamide (the product of step 18.) and 12-oxirane were mixed, sealed in THF containing chloroacetonitrile, heated at 80°C and stirred for 2 days until the target product was detected by MALDI-TOF mass spectrometry. The crude product was purified by silica gel chromatography, and the product was finally confirmed by nuclear magnetic resonance.
20.OT13-6的合成
20. Synthesis of OT13-6
OT13-6同步骤19.描述的合成方法相同，1-(十二烷基氨基)-N-{2-[4-(4-氧亚基-3,5-二氮杂十七烷-1-基)哌嗪-1-基]乙基}甲烷酰胺(步骤18.的产物)和环氧12烷混合，在含有氯乙腈的THF中密封80℃加热持续搅拌反应2天，直至MALDI-TOF质谱检测到目标产物。粗产物经由硅胶色谱纯化，产物最终由核磁确认。OT13-6 is the same as the synthesis method described in step 19. 1-(Dodecylamino)-N-{2-[4-(4-oxyylidene-3,5-diazaheptadecane-1-yl)piperazine-1-yl]ethyl}methaneamide (the product of step 18.) and 12-oxirane are mixed, sealed in THF containing chloroacetonitrile, heated at 80°C and stirred for 2 days until the target product is detected by MALDI-TOF mass spectrometry. The crude product is purified by silica gel chromatography, and the product is finally confirmed by nuclear magnetic resonance.
21.OT13-7的合成
21. Synthesis of OT13-7
OT13-7同步骤19.描述的合成方法相同，反应物以含有不饱和链2-[(6E)-十二-6-烯基]氧杂环丙烷替换步骤18.中的环氧12烷，在含有氯乙腈的THF中密封80℃加热持续搅拌反应2 天，直至MALDI-TOF质谱检测到目标产物。粗产物经由硅胶色谱纯化，产物最终由核磁确认。OT13-7 is the same as the synthesis method described in step 19. The reactant is replaced with an unsaturated chain 2-[(6E)-dode-6-enyl] oxirane in step 18. The mixture is sealed in THF containing chloroacetonitrile and heated at 80°C for 2 hours with continuous stirring. The reaction was continued for 14 days until the target product was detected by MALDI-TOF mass spectrometry. The crude product was purified by silica gel chromatography and the product was finally confirmed by NMR.
OT13-8的合成步骤Synthesis steps of OT13-8
22. 2-{[2-(4-{2-[(2-氨基乙基)氧基]乙基}哌嗪-1-基)乙基]氧基}乙-1-胺(2,2’-((piperazine-1,4-diylbis(ethane-2,1-diyl))bis(oxy))bis(ethan-1-amine))的合成
22. Synthesis of 2-{[2-(4-{2-[(2-aminoethyl)oxy]ethyl}piperazin-1-yl)ethyl]oxy}ethan-1-amine (2,2'-((piperazine-1,4-diylbis(ethane-2,1-diyl))bis(oxy))bis(ethan-1-amine)
1,4-双(2-羟乙基)哌嗪溶解于DMF和乙腈的混合体系，加入2-溴乙胺氢溴酸盐在酸性条件下并再溴化钾的催化下，50-100℃反应六个小时。1,4-bis(2-hydroxyethyl)piperazine is dissolved in a mixed system of DMF and acetonitrile, 2-bromoethylamine hydrobromide is added, and the mixture is reacted at 50-100°C for six hours under acidic conditions and catalyzed by potassium bromide.
23. 1-(十二烷基氨基)-N-[2-({2-[4-(7-氧亚基-6-氮杂-3-氧杂二十烷-1-基)哌嗪-1-基]乙基}氧基)乙基]甲烷酰胺(N-(2-(2-(4-(2-(2-(3-dodecylureido)ethoxy)ethyl)piperazin-1-yl)ethoxy)ethyl)tetradecanamide)的合成
23. Synthesis of 1-(dodecylamino)-N-[2-({2-[4-(7-oxydeoxy-6-aza-3-oxaicosane-1-yl)piperazin-1-yl]ethyl}oxy)ethyl]methaneamide (N-(2-(2-(4-(2-(2-(3-dodecylureido)ethoxy)ethyl)piperazin-1-yl)ethoxy)ethyl)tetradecanamide)
2-{[2-(4-{2-[(2-氨基乙基)氧基]乙基}哌嗪-1-基)乙基]氧基}乙-1-胺(步骤22.的产物)在CDI的催化下与十二烷-1-胺在THF下加热至70℃，通入新戊烷，震荡反应2小时。产物由硅胶色谱柱纯化，得到浅黄色固体。2-{[2-(4-{2-[(2-aminoethyl)oxy]ethyl}piperazin-1-yl)ethyl]oxy}ethan-1-amine (the product of step 22.) was heated to 70°C in THF with dodecane under the catalysis of CDI, and new pentane was introduced and shaken for 2 hours. The product was purified by silica gel column chromatography to obtain a light yellow solid.
24.OT13-8的合成
24. Synthesis of OT13-8
1-(十二烷基氨基)-N-[2-({2-[4-(7-氧亚基-6-氮杂-3-氧杂二十烷-1-基)哌嗪-1-基]乙基}氧基)乙基]甲烷酰胺(步骤23.的产物)和环氧12烷混合，在含有氯乙腈的THF中密封80℃加热持续搅拌反应2天，直至MALDI-TOF质谱检测到目标产物。粗产物经由硅胶色谱纯化，产物最终由核磁确认。1-(Dodecylamino)-N-[2-({2-[4-(7-oxyylidene-6-aza-3-oxaicosane-1-yl)piperazin-1-yl]ethyl}oxy)ethyl]methaneamide (the product of step 23.) was mixed with 12-oxirane oxide, and heated at 80°C in THF containing chloroacetonitrile and stirred for 2 days until the target product was detected by MALDI-TOF mass spectrometry. The crude product was purified by silica gel chromatography, and the product was finally confirmed by nuclear magnetic resonance.
OT13-9的合成步骤Synthesis steps of OT13-9
25.{[(9S)-12-(甲氧基羰基)-9-({[(2-甲基丙-2-基)氧基]羰基}氨基)-3,10-二氧亚基-1-苯基-4,11-二氮杂-2-氧杂十六烷-16-基]氨基}甲烷酸苄基酯(methyl N6-((benzyloxy)carbonyl)-N2-(N6-((benzyloxy)carbonyl)-N2-(tert-butoxycarbonyl)lysyl)lysinate)的合成
25. Synthesis of benzyl {[(9S)-12-(methoxycarbonyl)-9-({[(2-methylpropan-2-yl)oxy]carbonyl}amino)-3,10-dioxyylidene-1-phenyl-4,11-diaza-2-oxahexadecan-16-yl]amino}methanoate (methyl N 6 -((benzyloxy)carbonyl)-N 2 -(N 6 -((benzyloxy)carbonyl)-N 2 -(tert-butoxycarbonyl)lysyl)lysinate)
6-{[(苄基氧基)羰基]氨基}-2-({[(2-甲基丙-2-基)氧基]羰基}氨基)己酸(N6-((benzyloxy)carbonyl)-N2-(tert-butoxycarbonyl)lysine)与[(5-氨基-6-甲氧基-6-氧亚基己基)氨基]甲烷酸苄基酯(methyl N6-((benzyloxy)carbonyl)lysinate)混合，在EDCI，HOBt和TEA的催化下，乙腈作为溶剂25℃持续搅拌反应12小时。反应结束后将溶剂减压蒸馏，剩余粗产物在硅胶色谱柱下纯化。6-{[(benzyloxy)carbonyl]amino}-2-({[(2-methylprop-2-yl)oxy]carbonyl}amino)hexanoic acid (N 6 -((benzyloxy)carbonyl)-N 2 -(tert-butoxycarbonyl)lysine) and [(5-amino-6-methoxy-6-oxyylidenehexyl)amino]methanoic acid benzyl ester (methyl N 6 -((benzyloxy)carbonyl)lysinate) were mixed, and the mixture was stirred for 12 hours at 25°C in the presence of EDCI, HOBt and TEA, and acetonitrile was used as solvent. After the reaction, the solvent was distilled off under reduced pressure, and the remaining crude product was purified by silica gel column chromatography.
26.{[9-氨基-12-(甲氧基羰基)-3,10-二氧亚基-1-苯基-4,11-二氮杂-2-氧杂十六烷-16-基]氨基}甲烷酸苄基酯(methyl N6-((benzyloxy)carbonyl)-N2-(N6-((benzyloxy)carbonyl)lysyl)lysinate)的合成
26. Synthesis of methyl N6-((benzyloxy)carbonyl)-N2-(N6-((benzyloxy)carbonyl)lysyl)lysinate
{[(9S)-12-(甲氧基羰基)-9-({[(2-甲基丙-2-基)氧基]羰基}氨基)-3,10-二氧亚基-1-苯基-4,11-二氮杂-2-氧杂十六烷-16-基]氨基}甲烷酸苄基酯(步骤25.的产物)与1,2-二噁烷在25℃下反应5小时，滴加盐酸去除Boc基团。Benzyl {[(9S)-12-(methoxycarbonyl)-9-({[(2-methylpropan-2-yl)oxy]carbonyl}amino)-3,10-dioxyylidene-1-phenyl-4,11-diaza-2-oxahexadecan-16-yl]amino}methanoate (the product of step 25.) was reacted with 1,2-dioxane at 25° C. for 5 hours, and hydrochloric acid was added dropwise to remove the Boc group.
27. 2-[(9-氨基-3,10-二氧亚基-1-苯基-4-氮杂-2-氧杂癸-10-基)氨基]-6-{[(苄基氧基)羰基]氨基}己酸(N6-((benzyloxy)carbonyl)-N2-(N6-((benzyloxy)carbonyl)lysyl)lysine)的合成
27. Synthesis of 2-[(9-amino-3,10-dioxyylidene-1-phenyl-4-aza-2-oxadecan-10-yl)amino]-6-{[(benzyloxy)carbonyl]amino}hexanoic acid (N6-((benzyloxy)carbonyl)-N2-(N6-((benzyloxy)carbonyl)lysyl)lysine)
将{[9-氨基-12-(甲氧基羰基)-3,10-二氧亚基-1-苯基-4,11-二氮杂-2-氧杂十六烷-16-基]氨基}甲烷酸苄基酯(步骤26.的产物)溶解于甲醇，碱性环境(氢氧化钠调节)下25℃反应12小时。产物用水洗涤并真空干燥。Benzyl {[9-amino-12-(methoxycarbonyl)-3,10-dioxyylidene-1-phenyl-4,11-diaza-2-oxahexadecan-16-yl]amino}methane (the product of step 26) was dissolved in methanol and reacted at 25° C. for 12 hours in an alkaline environment (adjusted by sodium hydroxide). The product was washed with water and dried in vacuo.
28.({4-[3,6-二氧亚基-5-(3-氧亚基-1-苯基-4-氮杂-2-氧杂辛-8-基)哌嗪-2-基]丁基}氨基)甲烷酸苄基酯(dibenzyl((3,6-dixopiperazine-2,5-diyl)bis(butane-4,1-diyl))dicarbamate)的 合成
28. ({4-[3,6-dioxo-5-(3-oxo-1-phenyl-4-aza-2-oxaoctane-8-yl)piperazin-2-yl]butyl}amino)methanoic acid benzyl ester (dibenzyl((3,6-dixopiperazine-2,5-diyl)bis(butane-4,1-diyl))dicarbamate) synthesis
2-[(9-氨基-3,10-二氧亚基-1-苯基-4-氮杂-2-氧杂癸-10-基)氨基]-6-{[(苄基氧基)羰基]氨基}己酸(步骤27.的产物)在乙腈中溶解，在EDCI，HOBt和TEA的催化下持续搅拌反应12小时进行环酰化。加入水和乙酸乙酯停止反应。分离有机层，用硫酸镁进行干燥，并将剩余溶剂减压蒸馏，产物由硅胶色谱柱纯化。2-[(9-amino-3,10-dioxyylidene-1-phenyl-4-aza-2-oxadecan-10-yl)amino]-6-{[(benzyloxy)carbonyl]amino}hexanoic acid (the product of step 27.) was dissolved in acetonitrile and stirred for 12 hours under the catalysis of EDCI, HOBt and TEA for cycloacylation. Water and ethyl acetate were added to stop the reaction. The organic layer was separated and dried over magnesium sulfate, and the remaining solvent was distilled under reduced pressure. The product was purified by silica gel column chromatography.
29. 3,6-双(4-氨基丁基)哌嗪-2,5-二酮(3,6-bis(4-aminobutyl)piperazine-2,5-dione)的合成
29. Synthesis of 3,6-bis(4-aminobutyl)piperazine-2,5-dione
干燥圆底烧瓶里通氢气，将({4-[3,6-二氧亚基-5-(3-氧亚基-1-苯基-4-氮杂-2-氧杂辛-8-基)哌嗪-2-基]丁基}氨基)甲烷酸苄基酯(步骤28.的产物)加入含有乙酸的二氯甲烷溶剂中，在Pd或者C的催化下，165℃油浴反应12小时。该反应需维持在500psi压力下，通过不锈钢压力容器实现。产物过滤后旋蒸浓缩，得到浅黄色油状物。A dry round-bottom flask was filled with hydrogen, and ({4-[3,6-dioxy-5-(3-oxy-1-phenyl-4-aza-2-oxaoctane-8-yl)piperazine-2-yl]butyl}amino)benzyl methane ester (the product of step 28.) was added to a dichloromethane solvent containing acetic acid, and reacted in an oil bath at 165°C for 12 hours under the catalysis of Pd or C. The reaction was maintained at a pressure of 500 psi and was achieved by a stainless steel pressure vessel. The product was filtered and concentrated by rotary evaporation to obtain a light yellow oil.
30. 3,6-双{4-[双(2-氨基乙基)氨基]丁基}哌嗪-2,5-二酮(3,6-bis(4-(bis(2-aminoethyl)amino)butyl)piperazine-2,5-dione)的合成
30. Synthesis of 3,6-bis(4-(bis(2-aminoethyl)amino)butyl)piperazine-2,5-dione
3,6-双(4-氨基丁基)哌嗪-2,5-二酮(步骤29.的产物)和N-[二甲基(2-甲基丙-2-基)甲硅基]甲烷酰胺在含有TEA的盐酸体系下室温振摇反应17小时，通过NaBH(OAc)3还原TBS基团。产物过滤后旋蒸浓缩，得到浅黄色油状物。3,6-bis(4-aminobutyl)piperazine-2,5-dione (the product of step 29) and N-[dimethyl(2-methylprop-2-yl)silyl]methaneamide were shaken at room temperature for 17 hours in a hydrochloric acid system containing TEA, and the TBS group was reduced by NaBH(OAc) 3. The product was filtered and concentrated by rotary evaporation to obtain a light yellow oil.
31.OT13-9的合成
31. Synthesis of OT13-9
3,6-双{4-[双(2-氨基乙基)氨基]丁基}哌嗪-2,5-二酮(步骤30.的产物)在CDI的催化下与(8E)-十六-8-烯-1-胺在THF下加热至70℃，通入新戊烷，震荡反应2小时。产物由硅胶色谱柱纯化，得到浅黄色固体。3,6-bis{4-[bis(2-aminoethyl)amino]butyl}piperazine-2,5-dione (the product of step 30) was heated to 70°C in THF with (8E)-hexa-8-en-1-amine under the catalysis of CDI, and neopentane was introduced and shaken for 2 hours. The product was purified by silica gel column chromatography to obtain a light yellow solid.
OT13-10的合成步骤Synthesis steps of OT13-10
32. 2-(十一-4-炔基氧基)四氢吡喃(2-(undec-4-yn-1-yloxy)tetrahydro-2H-pyran)的合成
32. Synthesis of 2-(undec-4-yn-1-yloxy)tetrahydro-2H-pyran
2-(戊-4-炔基氧基)四氢吡喃溶解于THF和HMPA体系，在聚合起始剂正丁基锂的作用下和1-溴己烷反应，生成产物，经色谱柱纯化。2-(Pent-4-ynyloxy)tetrahydropyran is dissolved in THF and HMPA system, reacted with 1-bromohexane under the action of polymerization initiator n-butyl lithium to generate a product, which is purified by chromatography column.
33. 2-{[(4Z)-十一-4-烯基]氧基}四氢吡喃((Z)-2-(undec-4-en-1-yloxy)tetrahydro-2H-pyran)的合成
33. Synthesis of 2-{[(4Z)-undec-4-enyl]oxy}tetrahydropyran ((Z)-2-(undec-4-en-1-yloxy)tetrahydro-2H-pyran)
步骤32.的产物在林德拉催化剂的催化下，乙醇反应体系进行氢化反应，生成2-{[(4Z)-十一-4-烯基]氧基}四氢吡喃，产物经由硅胶色谱柱粗纯。The product of step 32. is subjected to hydrogenation reaction in an ethanol reaction system under the catalysis of Lindela catalyst to generate 2-{[(4Z)-undec-4-enyl]oxy}tetrahydropyran, and the product is purified by silica gel column chromatography.
34.(4Z)-十一-4-烯-1-醇((Z)-undec-4-en-1-ol)的合成
34. Synthesis of (4Z)-undec-4-en-1-ol
對甲苯磺酸溶解于甲醇，与步骤33.的产物反应，生成(4Z)-十一-4-烯-1-醇，产物经有硅胶色谱柱粗纯。p-Toluenesulfonic acid is dissolved in methanol and reacted with the product of step 33 to generate (4Z)-undec-4-en-1-ol. The product is purified by silica gel column chromatography.
35.甲烷磺酸-(4Z)-十一-4-烯-1-基酯((Z)-undec-4-en-1-yl methanesulfonate)的合成
35. Synthesis of (4Z)-undec-4-en-1-yl methanesulfonate
室温下，将MsCl(甲磺酰氯)溶解于二氯甲烷中，在惰性气体条件下，将((4Z)-十一-4-烯-1-醇与甲磺酰氯在0-20℃反应温度及三乙胺催化下进行温和搅拌。用蒸馏水终止反应，并洗涤总有机层，再用饱和卤盐洗涤，干燥，浓缩。用硅胶柱色谱纯化直至纯度大于85％。 At room temperature, MsCl (methanesulfonyl chloride) was dissolved in dichloromethane. Under inert gas conditions, ((4Z)-undec-4-ene-1-ol and methanesulfonyl chloride were gently stirred at a reaction temperature of 0-20°C and catalyzed by triethylamine. The reaction was terminated with distilled water, and the total organic layer was washed with saturated halide, dried, and concentrated. Purification was performed by silica gel column chromatography until the purity was greater than 85%.
36.(4Z)-1-叠氮十一-4-烯((Z)-1-azidoundec-4-ene)的合成
36. Synthesis of (4Z)-1-azidoundec-4-ene
在DMSO体系下将步骤35.的产物同叠氮化钠进行反应，生成的产物经硅胶色谱柱纯化。The product of step 35. is reacted with sodium azide in the presence of DMSO, and the resulting product is purified by silica gel column chromatography.
37.(4Z)-十一-4-烯-1-胺((Z)-undec-4-en-1-amine)的合成
37. Synthesis of (4Z)-undec-4-en-1-amine
将步骤36.的产物在THF溶剂中通过水合三苯基膦催化进行氢化取代还原。产物通过硅胶色谱柱进行纯化并备用The product of step 36. is subjected to hydrogenation substitution reduction in THF solvent by catalysis of triphenylphosphine hydrate. The product is purified by silica gel chromatography and used as
38.[(2,2-二甲基-4,10-二氧亚基-5-氮杂-3-氧杂十四烷-14-基)氨基]甲烷酸-2-甲基丙-2-基酯(di-tert-butyl(5-oxononane-1,9-diyl)dicarbamate)的合成
38. Synthesis of 2-methylpropane-2-yl[(2,2-dimethyl-4,10-dioxy-5-aza-3-oxatetradec-14-yl)amino]methane (di-tert-butyl(5-oxononane-1,9-diyl)dicarbamate)
[(4-溴丁基)氨基]甲烷酸-2-甲基丙-2-基酯在含有镁粉和碘的THF反应溶剂中和2-氧亚基六氢吡啶-1-甲酸-2-甲基丙-2-基酯进行反映，产物经硅胶色谱柱纯化。[(4-Bromobutyl)amino]methane acid-2-methylpropan-2-yl ester is reacted with 2-oxo-pyridinium-1-carboxylic acid-2-methylpropan-2-yl ester in a THF reaction solvent containing magnesium powder and iodine, and the product is purified by silica gel column chromatography.
39.[(10-氨基-2,2-二甲基-4-氧亚基-5-氮杂-3-氧杂十四烷-14-基)氨基]甲烷酸-2-甲基丙-2-基酯(di-tert-butyl(5-aminononane-1,9-diyl)dicarbamate)的合成
39. Synthesis of 2-methylpropane-2-yl[(10-amino-2,2-dimethyl-4-oxyylidene-5-aza-3-oxatetradecane-14-yl)amino]methane (di-tert-butyl(5-aminononane-1,9-diyl)dicarbamate)
在乙醇和乙酸铵混合反应体系中，室温下加入氰基硼氢化钠，持续搅拌超过2小时。产物经矽鈣石过滤，并且用热乙酸乙酯洗涤。Sodium cyanoborohydride was added to a mixed reaction system of ethanol and ammonium acetate at room temperature and stirred for more than 2 hours. The product was filtered through silica and washed with hot ethyl acetate.
40.{[8-(2,2-二甲基-4-氧亚基-5-氮杂-3-氧杂壬-9-基)-2-甲基-6-氧亚基-2,5,7-三氮杂十二烷-12-基]氨基}甲烷酸-2-甲基丙-2-基酯(di-tert-butyl(5-(3-(2-(dimethylamino)ethyl)ureido)nonane-1,9-diyl)dicarbamate)的合成
40. Synthesis of 2-methylpropane-2-yl{[8-(2,2-dimethyl-4-oxyde-5-aza-3-oxanonan-9-yl)-2-methyl-6-oxyde-2,5,7-triazadodec-12-yl]amino}methanoate (di-tert-butyl(5-(3-(2-(dimethylamino)ethyl)ureido)nonane-1,9-diyl)dicarbamate)
[(10-氨基-2,2-二甲基-4-氧亚基-5-氮杂-3-氧杂十四烷-14-基)氨基]甲烷酸-2-甲基丙-2-基酯(步骤39.的产物)在CDI的催化下与2-(二甲基氨基)乙-1-胺在THF下加热至70℃，通入新戊烷，震荡反应2小时。产物由硅胶色谱柱纯化，得到产物。 [(10-amino-2,2-dimethyl-4-oxyylidene-5-aza-3-oxatetradecane-14-yl)amino]methanoic acid-2-methylpropan-2-yl ester (the product of step 39.) was heated to 70° C. in THF with 2-(dimethylamino)ethan-1-amine under the catalysis of CDI, and new pentane was introduced and shaken for 2 hours. The product was purified by silica gel column chromatography to obtain the product.
41.OT13-10的合成
41. Synthesis of OT13-10
将步骤40.的产物在乙酸乙酯/盐酸混合体系下进行脱Boc保护，产物利用硅胶色谱柱纯化，得到产物随即同(4Z)-十一-4-烯-1-胺(步骤37.产物)在THF下加热至70℃，通入新戊烷，震荡反应2小时。产物由硅胶色谱柱纯化，得到OT13-10。The product of step 40. was deprotected from Boc in a mixed system of ethyl acetate/hydrochloric acid, and the product was purified by silica gel chromatography to obtain the product, which was then heated to 70° C. with (4Z)-undec-4-en-1-amine (product of step 37.) in THF, and neopentane was introduced and shaken for 2 hours. The product was purified by silica gel chromatography to obtain OT13-10.
OT13-11的合成步骤Synthesis steps of OT13-11
42.[(1-苯基-7-氮杂-2-氧杂十三烷-13-基)氨基]甲烷酸-2-甲基丙-2-基酯(tert-butyl(6-((4-(benzyloxy)butyl)amino)hexyl)carbamate)的合成
42. Synthesis of 2-methylpropan-2-yl[(1-phenyl-7-aza-2-oxatridec-13-yl)amino]methane ester (tert-butyl(6-((4-(benzyloxy)butyl)amino)hexyl)carbamate)
[(6-氨基己基)氨基]甲烷酸-2-甲基丙-2-基酯同{[(4-溴丁基)氧基]甲基}苯在含有碳酸钾的乙腈溶剂条件下进行反应，生成产物。[(6-aminohexyl)amino]methanoic acid-2-methylpropan-2-yl ester reacts with {[(4-bromobutyl)oxy]methyl}benzene in an acetonitrile solvent containing potassium carbonate to produce a product.
43.({12-[4-(苄基氧基)丁基]-2,2-二甲基-4-氧亚基-5,12-二氮杂-3-氧杂十八烷-18-基}氨基)甲烷酸-2-甲基丙-2-基酯(di-tert-butyl(((4-(benzyloxy)butyl)azanediyl)bis(hexane-6,1-diyl))dicarbamate)的合成
43. Synthesis of 2-methylpropanediyl (di-tert-butyl (((4-(benzyloxy)butyl)azanediyl)bis(hexane-6,1-diyl))dicarbamate)
在甲醇中，室温下加入氰基硼氢化钠，步骤42.的产物同[(5-甲酰基戊基)氨基]甲烷酸-2-甲基丙-2-基酯进行反应，持续搅拌超过2小时。产物经矽鈣石过滤，并且用热乙酸乙酯洗涤。The product of step 42. was reacted with 2-methylpropan-2-yl [(5-formylpentyl)amino]methanoate in methanol at room temperature by adding sodium cyanoborohydride. The stirring was continued for more than 2 hours. The product was filtered through silica and washed with hot ethyl acetate.
44. 6-(13-氨基-1-苯基-7-氮杂-2-氧杂十三烷-7-基)己-1-胺(N1-(6-aminohexyl)-N1-(4-(benzyloxy)butyl)hexane-1,6-diamine)的合成
44. Synthesis of 6-(13-amino-1-phenyl-7-aza-2-oxatridecan-7-yl)hexane-1-amine (N1-(6-aminohexyl)-N1-(4-(benzyloxy)butyl)hexane-1,6-diamine)
将步骤43.的产物在乙酸乙酯/盐酸混合体系下进行脱Boc保护；另外庚-1-胺和1-碘己烷在有碳酸钾的乙腈溶剂条件下进行反应，生成产物己基(辛基)胺，经硅胶色谱柱纯化后待下一步反应使用。The product of step 43. is deprotected from Boc in a mixed system of ethyl acetate/hydrochloric acid; in addition, heptyl-1-amine and 1-iodohexane are reacted in an acetonitrile solvent containing potassium carbonate to generate the product hexyl (octyl) amine, which is purified by a silica gel column and used in the next reaction.
45. 1-({7-[4-(苄基氧基)丁基]-15-氧亚基-16-戊基-7,14,16-三氮杂二十四烷-1-基}氨基)-N-辛基-N-戊基甲烷酰胺(1,1'-(((4-(benzyloxy)butyl)azanediyl)bis(hexane-6,1-diyl))bis(3-hexyl-3-octylurea))的合成
45. Synthesis of 1-({7-[4-(benzyloxy)butyl]-15-oxydeca-16-pentyl-7,14,16-triazatetracosane-1-yl}amino)-N-octyl-N-pentylmethaneamide (1,1'-(((4-(benzyloxy)butyl)azanediyl)bis(hexane-6,1-diyl))bis(3-hexyl-3-octylurea))
步骤43.的产物脱Boc后得到的产物在CDI的催化下与己基(辛基)胺在THF下加热至70℃，通入新戊烷，震荡反应2小时。经硅胶色谱柱纯化，得到产物。The product obtained after de-Bocation of the product in step 43. is heated to 70° C. in THF with hexyl(octyl)amine under the catalysis of CDI, and neopentane is introduced to react with shaking for 2 hours. The product is purified by silica gel column chromatography to obtain the product.
46.OT13-11的合成
46. Synthesis of OT13-11
干燥圆底烧瓶里通氢气，将1-({7-[4-(苄基氧基)丁基]-15-氧亚基-16-戊基-7,14,16-三氮杂二十四烷-1-基}氨基)-N-辛基-N-戊基甲烷酰胺(步骤45.产物)加入乙醇溶剂中，在Pd的催化下，165℃油浴反应12小时。该反应需维持在500psi压力下，通过不锈钢压力容器实现。过滤后旋蒸浓缩，得到OT13-11。A dry round-bottom flask was filled with hydrogen, and 1-({7-[4-(benzyloxy)butyl]-15-oxyylidene-16-pentyl-7,14,16-triazatetracosane-1-yl}amino)-N-octyl-N-pentylmethaneamide (product of step 45) was added to an ethanol solvent, and reacted in an oil bath at 165°C for 12 hours under the catalysis of Pd. The reaction was maintained at a pressure of 500 psi and was achieved by a stainless steel pressure vessel. After filtration, the mixture was concentrated by rotary evaporation to obtain OT13-11.
OT13-12的合成步骤Synthesis steps of OT13-12
47.[(1-苯基-5-氮杂-2-氧杂癸-10-基)氨基]甲烷酸-2-甲基丙-2-基酯(tert-butyl(5-((2-(benzyloxy)ethyl)amino)pentyl)carbamate)的合成
47. Synthesis of 2-methylpropan-2-yl[(1-phenyl-5-aza-2-oxadecan-10-yl)amino]methaneate (tert-butyl(5-((2-(benzyloxy)ethyl)amino)pentyl)carbamate)
[(5-溴戊基)氨基]甲烷酸-2-甲基丙-2-基酯同2-(苄基氧基)乙-1-胺常温下在含有碳酸钾的乙腈溶剂中进行反应，生成产物，经硅胶色谱柱纯化。[(5-Bromopentyl)amino]methane acid-2-methylprop-2-yl ester reacts with 2-(benzyloxy)ethyl-1-amine in an acetonitrile solvent containing potassium carbonate at room temperature to generate a product, which is purified by silica gel column chromatography.
48.({11-[2-(苄基氧基)乙基]-2,2-二甲基-4-氧亚基-5,11-二氮杂-3-氧杂十五烷-15-基}氨基)甲烷酸苄基酯(benzyl(4-((2-(benzyloxy)ethyl)(5-((tert butoxycarbonyl)amino)pentyl)amino)butyl)carbamate)的合成
48. Synthesis of benzyl(4-((2-(benzyloxy)ethyl)(5-((tert butoxycarbonyl)amino)pentyl)amino)butyl)carbamate
在步骤47.的产物中，室温下加入[(3-甲酰基丙基)氨基]甲烷酸苄基酯，在氰基硼氢化钠的催化下，持续搅拌超过2小时。产物经矽鈣石过滤，并且用热乙酸乙酯洗涤备用。To the product of step 47., benzyl [(3-formylpropyl)amino]methanoate was added at room temperature, and the mixture was stirred for more than 2 hours under the catalysis of sodium cyanoborohydride. The product was filtered through silica and washed with hot ethyl acetate for later use.
49.{[5-(5-氨基戊基)-1-苯基-5-氮杂-2-氧杂壬-9-基]氨基}甲烷酸苄基酯(benzyl(4-((5-aminopentyl)(2-(benzyloxy)ethyl)amino)butyl)carbamate)的合成
49. Synthesis of benzyl (4-((5-aminopentyl)(2-(benzyloxy)ethyl)amino)butyl)carbamate
将步骤48.的产物在乙酸乙酯/盐酸混合体系下进行脱Boc保护，产物经硅胶色谱柱纯化后待下一步反应使用。The product of step 48 was de-Boc protected in an ethyl acetate/hydrochloric acid mixture system, and the product was purified by silica gel column chromatography and used in the next reaction.
50. 2-辛基癸酸(2-octyldecanoic acid)的合成
50. Synthesis of 2-octyldecanoic acid
1-溴辛烷在THF溶剂中，在氢化钠的催化下常温同癸酸在持续搅拌下反应两个小时，生成2-辛基癸酸，产物经硅胶色谱柱纯化备用。1-Bromooctane is reacted with decanoic acid in THF solvent under the catalysis of sodium hydride at room temperature under continuous stirring for two hours to generate 2-octyldecanoic acid, and the product is purified by silica gel chromatography column for use.
51. 2-辛基癸-1-醇(2-octyldecan-1-ol)的合成
51. Synthesis of 2-octyldecan-1-ol
2-辛基癸酸溶解在THF溶剂里，室温下持续搅拌2小时，通过LaN催化还原生成脂质纳米颗粒的制备和表征2-辛基癸-1-醇，产物经硅胶色谱柱纯化。2-Octyldecanoic acid was dissolved in THF solvent and stirred continuously at room temperature for 2 h. 2-Octyldecan-1-ol was generated by LaN-catalyzed reduction and the product was purified by silica gel column chromatography.
52. 2-(2-辛基癸基)异吲哚-1,3-二酮(2-(2-octyldecyl)isoindoline-1,3-dione)的合成
52. Synthesis of 2-(2-octyldecyl)isoindoline-1,3-dione
室温下，将Ms2O(甲磺酸酐)溶解于含有TEA的二氯甲烷中，在惰性气体条件下，与2-辛基癸-1-醇(步骤51.的产物)在0-25℃反应温度下进行温和搅拌，生成产物。用蒸馏水终止反应，并洗涤总有机层，再用饱和卤盐洗涤，干燥，浓缩。用硅胶柱色谱纯化直至纯度大于85％。之后邻苯二甲酰亚胺钾盐溶解于DMF，并在室温下加入叠氮化钠。在70℃反应温度下持续搅拌2小时，减压蒸馏。加入二氯甲烷萃取，并用水洗涤，经无水硫酸钠干燥后得到粗产物，并用色谱柱进行分离纯化。At room temperature, Ms 2 O (methanesulfonic anhydride) is dissolved in dichloromethane containing TEA, and gently stirred with 2-octyldecan-1-ol (the product of step 51.) at a reaction temperature of 0-25°C under inert gas conditions to generate a product. The reaction is terminated with distilled water, and the total organic layer is washed, then washed with saturated halide, dried, and concentrated. Purification is performed by silica gel column chromatography until the purity is greater than 85%. Thereafter, potassium phthalimide is dissolved in DMF, and sodium azide is added at room temperature. Stirring is continued at a reaction temperature of 70°C for 2 hours, and distillation is carried out under reduced pressure. Dichloromethane is added for extraction, and the mixture is washed with water. The crude product is obtained after drying over anhydrous sodium sulfate, and separated and purified by a chromatographic column.
53. 2-辛基癸-1-胺(2-octyldecan-1-amine)的合成
53. Synthesis of 2-octyldecan-1-amine
步骤52.的产物进行Gabriel伯胺反应，反应体系为乙醇，在80℃温度下同水合肼作用，生成2-辛基癸-1-胺，产物经硅胶色谱柱纯化。 The product of step 52 is subjected to Gabriel primary amine reaction, the reaction system is ethanol, and it reacts with hydrazine hydrate at 80° C. to generate 2-octyldec-1-amine, and the product is purified by silica gel column chromatography.
54.({5-[2-(苄基氧基)乙基]-15-辛基-12-氧亚基-5,11,13-三氮杂二十三烷-1-基}氨基)甲烷酸苄基酯(benzyl(4-((2-(benzyloxy)ethyl)(5-(3-(2-octyldecyl)ureido)pentyl)amino)butyl)carbamate)的合成
54. Synthesis of benzyl(4-((2-(benzyloxy)ethyl)(5-(3-(2-octyldecyl)ureido)pentyl)amino)butyl)carbamate
步骤53.得到的产物2-辛基癸-1-胺和步骤49.的产物在THF下加热至70℃，通入新戊烷，震荡反应2小时，得到产物并经由硅胶色谱柱纯化。The product 2-octyldec-1-amine obtained in step 53 and the product in step 49 are heated to 70° C. in THF, and new pentane is introduced. The mixture is shaken for 2 hours to obtain the product, which is then purified via silica gel chromatography.
55. 1-({5-[(4-氨基丁基)(2-羟基乙基)氨基]戊基}氨基)-N-(2-辛基癸基)甲烷酰胺(1-(5-((4-aminobutyl)(2-hydroxyethyl)amino)pentyl)-3-(2-octyldecyl)urea)的合成
55. Synthesis of 1-({5-[(4-aminobutyl)(2-hydroxyethyl)amino]pentyl}amino)-N-(2-octyldecyl)methaneamide (1-(5-((4-aminobutyl)(2-hydroxyethyl)amino)pentyl)-3-(2-octyldecyl)urea)
干燥圆底烧瓶里通氢气，将步骤54.的产物加入含有乙酸的二氯甲烷溶剂中，在Pd或者C的催化下，常温反应2h，进行脱苯还原。该反应需维持在500psi压力下，通过不锈钢压力容器实现。产物过滤后旋蒸浓缩，并通过硅胶色谱柱纯化。A dry round-bottom flask is filled with hydrogen, and the product of step 54 is added to a dichloromethane solvent containing acetic acid, and reacted at room temperature for 2 hours under the catalysis of Pd or C to perform debenzenization reduction. The reaction needs to be maintained at a pressure of 500 psi and is achieved in a stainless steel pressure vessel. The product is filtered, concentrated by rotary evaporation, and purified by silica gel chromatography.
56.OT13-12的合成
56. Synthesis of OT13-12
步骤55.得到的产物与十一烷-1-胺在THF中在70℃下，通入新戊烷，震荡反应2小时，得到OT13-12，产物经由硅胶色谱柱纯化。Step 55. The obtained product was reacted with undecane-1-amine in THF at 70°C, and neopentane was introduced, and the reaction was shaken for 2 hours to obtain OT13-12. The product was purified by silica gel chromatography.
实施例2Example 2
脂质纳米颗粒的制备和表征Preparation and characterization of lipid nanoparticles
通过充分混合有机相和水相，制备LNP纳米颗粒，实现基因编辑工具或其它核酸药物的有效包埋。By fully mixing the organic phase and the aqueous phase, LNP nanoparticles are prepared to achieve effective encapsulation of gene editing tools or other nucleic acid drugs.
其中，有机相：含有至少一种可电离脂质(OT13-1～OT13-9)、至少一种支撑脂质、至少一种双亲性嵌段共聚物以及胆固醇，由可以与水互溶的有机溶剂溶解。所述有机溶剂优选自乙醇、乙腈、丙酮等。本实施例中采用OT13-3,DSPC，PEG-DMG,以及胆固醇，其中四组分 的摩尔比是50:10:1.5:38.5；The organic phase contains at least one ionizable lipid (OT13-1 to OT13-9), at least one supporting lipid, at least one amphiphilic block copolymer and cholesterol, and is dissolved in an organic solvent that is miscible with water. The organic solvent is preferably selected from ethanol, acetonitrile, acetone, etc. In this embodiment, OT13-3, DSPC, PEG-DMG, and cholesterol are used, wherein the four components The molar ratio is 50:10:1.5:38.5;
水相：基因编辑工具的水溶液，其中，核酸物质采用酵母提取RNA作为模版RNA(麦克林，log.R822593)，含量为0.5-50％(w/v)(本实施例优选含量为40％(w/v))，pH为约3.0至约7.0(本实施例优选pH为约4.0)。水相盐溶液可选自：柠檬酸缓冲液、磷酸盐缓冲液、Tris-HCl缓冲液体系；其中，本实施例采用的是柠檬酸缓冲液。Aqueous phase: an aqueous solution of a gene editing tool, wherein the nucleic acid material uses yeast extracted RNA as a template RNA (McLean, log. R822593), the content is 0.5-50% (w/v) (the preferred content in this embodiment is 40% (w/v)), and the pH is about 3.0 to about 7.0 (the preferred pH in this embodiment is about 4.0). The aqueous salt solution can be selected from: citric acid buffer, phosphate buffer, Tris-HCl buffer system; wherein, the citric acid buffer is used in this embodiment.
有机相与水相的混合可以通过微流控及撞击流反应器实现。基因编辑工具RNA的包埋效率可以通过调控体系N/P比例进行优化，N/P比例为1:1至9:1。The mixing of organic and aqueous phases can be achieved by microfluidics and impinging flow reactors. The embedding efficiency of gene editing tool RNA can be optimized by regulating the N/P ratio of the system, which ranges from 1:1 to 9:1.
通过本实施例的方式制备LNP，测试粒径，分散度(PDI)，以及包埋率(EE)，结果显示在下表中：
LNP was prepared by the method of this example, and the particle size, dispersion (PDI), and embedding efficiency (EE) were tested. The results are shown in the following table:
其中，包埋率表征方法：制备的LNP包埋率通过Quant-iTTMRNA Reagent and Kit试剂盒测定。具体检测方法：Among them, the embedding rate characterization method: the embedding rate of the prepared LNP is measured by Quant-iTTM RNA Reagent and Kit. Specific detection method:
溶液配置：用超纯水将适量的20×TE缓冲液稀释成1×，够当日实验用量即可。用1×的TE(大范围25-1000ng/ml RNA)按1:200的比例，(小范围1-50ng/ml RNA)按1:2000的比例稀释浓缩染料液。用箔金纸包裹或放置黑暗处避光保存。用1×TE缓冲液稀释TritonX-100至5％浓度备用。Solution preparation: Dilute an appropriate amount of 20×TE buffer to 1× with ultrapure water, enough for the experiment on the day. Dilute the concentrated dye solution with 1×TE (large range 25-1000ng/ml RNA) at a ratio of 1:200, (small range 1-50ng/ml RNA) at a ratio of 1:2000. Wrap it with foil or store it in a dark place away from light. Dilute TritonX-100 with 1×TE buffer to a concentration of 5% for later use.
样品处理：设置RNA浓度为0的超纯水作为空白背景，设置加TE缓冲液代替加5％TritonX-100的LNP作为游离RNA测定样，设置加5％TritonX-100的LNP作为总RNA测定样。取需测定的LNP样品及空白中加入等体积的5％TritonX-100、1×TE溶液，50～60℃孵育5～10min。孵育后的LNP样品用1×TE缓冲液稀释至10-400倍(根据样品组RNA浓度适当调整)。稀释后取100μl加入96孔板中，而后避光添加100μl稀释好的浓缩染料液，5分钟内测定荧光吸收度。测定条件为激发波长480nm，发射波长520nm。Sample processing: Set ultrapure water with RNA concentration of 0 as blank background, set LNP with TE buffer instead of 5% TritonX-100 as free RNA determination sample, set LNP with 5% TritonX-100 as total RNA determination sample. Take the LNP sample to be determined and the blank and add equal volumes of 5% TritonX-100 and 1×TE solution, incubate at 50-60°C for 5-10 minutes. The incubated LNP sample is diluted to 10-400 times with 1×TE buffer (appropriately adjusted according to the RNA concentration of the sample group). After dilution, take 100μl and add it to a 96-well plate, and then add 100μl of the diluted concentrated dye solution in the dark, and measure the fluorescence absorbance within 5 minutes. The determination conditions are excitation wavelength 480nm, emission wavelength 520nm.
计算方法：包埋率＝(总RNA吸光度-游离RNA吸光度)/总RNA吸光度×100％。Calculation method: embedding rate = (total RNA absorbance - free RNA absorbance) / total RNA absorbance × 100%.
实施例3Example 3
小鼠肝脏递送效果评价Evaluation of delivery effect in mouse liver
通过实施例2的方式制备LNP，其中可电离脂质为OT13-3，包埋的核酸为Luciferase mRNA(核苷酸序列如下表所示)。制备好的Luc-mRNA-LNP注射到6-8周的C57BL/6小鼠中，利用活体成像技术观察注射后6小时小鼠肝脏Lurciferase表达水平(图1)，以此评价 OT13脂质形成LNP在小鼠肝脏的递送效果。结果显示注射后6小时，包含可电离脂质OT13-3的LNP成功将Luc-mRNA递送到小鼠肝脏，并实现高效Luciferase表达。
LNP was prepared by the method of Example 2, wherein the ionizable lipid was OT13-3 and the embedded nucleic acid was Luciferase mRNA (nucleotide sequence is shown in the following table). The prepared Luc-mRNA-LNP was injected into 6-8 week old C57BL/6 mice, and the expression level of Lurciferase in the mouse liver was observed by in vivo imaging technology 6 hours after injection (Figure 1), so as to evaluate Delivery effect of OT13 lipid-formed LNP in mouse liver. The results showed that 6 hours after injection, LNP containing ionizable lipid OT13-3 successfully delivered Luc-mRNA to mouse liver and achieved efficient Luciferase expression.
成年Ai9转基因小鼠静脉注射肝脏转染效果评价Evaluation of the effect of intravenous liver transfection in adult Ai9 transgenic mice
采用实施例2的方式制备含OT13-3可电离脂质的LNP，同时包埋Cre mRNA(核苷酸序列如下表所示)的样品。注射到成年Ai9转基因小鼠中。注射后5天取出小鼠肝脏进行冷冻切片，并用激光共聚焦显微镜进行观察，结果显示包含OT13-3的LNP可以有效转染并编辑成年鼠肝脏(图2)。
The LNP containing OT13-3 ionizable lipids was prepared in the manner of Example 2, and a sample of Cre mRNA (nucleotide sequence is shown in the table below) was embedded at the same time. It was injected into adult Ai9 transgenic mice. Five days after injection, the mouse liver was removed for frozen sectioning and observed with a laser confocal microscope. The results showed that LNP containing OT13-3 can effectively transfect and edit the adult mouse liver (Figure 2).
活体内小鼠PCSK9基因编辑效果评价Evaluation of PCSK9 gene editing effects in mice in vivo
PCSK9是适用于分析功能性基因编辑工具递送至肝脏的目标基因，其在肝脏细胞内产生，分泌到血液循环中，因而可以应用于肝脏定向的脂质分子活体筛选。针对PCSK9的具有沉默效果的基因编辑工具EPIREG-mRNA及gRNA(其靶向序列的互补核苷酸序列如下表所示)通过实施例2的方法被共同包封到含有OT13-3的LNP中，以3mg/kg RNA浓度注射到野生型6-8周的C57BL/6小鼠体内，9天后应用试剂盒进行PCSK9检测(图3)。结果显示OT13-3形成LNP具有近80％表观基因编辑及沉默目标基因的效果。OT13-3形成LNP在体内表观遗传编辑药效可以对标阳性对照，该阳性对照应用商业化应用于临床的可电离脂质形成的LNP。此实施例的评价数据证明OT13系列分子可以作为核酸递送系统应用于肝脏靶向的基因治疗领域。
PCSK9 is a target gene suitable for analyzing the delivery of functional gene editing tools to the liver. It is produced in liver cells and secreted into the blood circulation, so it can be used for in vivo screening of liver-directed lipid molecules. EPIREG-mRNA and gRNA (the complementary nucleotide sequence of its targeting sequence is shown in the following table), a gene editing tool with a silencing effect for PCSK9, are co-encapsulated into LNPs containing OT13-3 by the method of Example 2, and injected into wild-type 6-8-week C57BL/6 mice at a concentration of 3 mg/kg RNA. After 9 days, the kit was used for PCSK9 detection (Figure 3). The results show that OT13-3 forms LNPs with nearly 80% epigenetic editing and silencing of target genes. The epigenetic editing efficacy of OT13-3-formed LNPs in vivo can be compared with a positive control, which is commercially applied to LNPs formed by ionizable lipids for clinical use. The evaluation data of this embodiment proves that the OT13 series of molecules can be used as a nucleic acid delivery system in the field of liver-targeted gene therapy.
本实施例的核酸序列：









The nucleic acid sequence of this example:









前述详细说明是以解释和举例的方式提供的，并非要限制所附权利要求的范围。目前本申请所列举的实施方式的多种变化对本领域普通技术人员来说是显而易见的，且保留在所附的权利要求和其等同方案的范围内。 The foregoing detailed description is provided by way of explanation and example, and is not intended to limit the scope of the appended claims. Various changes to the embodiments currently listed in this application are obvious to those of ordinary skill in the art and are retained within the scope of the appended claims and their equivalents.

Claims (47)

  1. 一种化合物，或其药学上可接受的盐，所述化合物包含阳离子提供基团以及氢键提供基团，且所述阳离子提供基团与氢键提供基团之间包含2至6个原子。A compound, or a pharmaceutically acceptable salt thereof, comprising a cation providing group and a hydrogen bond providing group, wherein the cation providing group and the hydrogen bond providing group contain 2 to 6 atoms.
  2. 如权利要求1所述的化合物，所述化合物为阳离子脂质化合物。The compound according to claim 1, wherein the compound is a cationic lipid compound.
  3. 如权利要求1-2中任一项所述的化合物，所述化合物为可电离的阳离子脂质化合物。The compound according to any one of claims 1 to 2, which is an ionizable cationic lipid compound.
  4. 如权利要求1-3中任一项所述的化合物，所述阳离子提供基团包含在pH为约7.0或更低的条件下带正电的基团。The compound of any one of claims 1 to 3, wherein the cation providing group comprises a group that is positively charged at a pH of about 7.0 or less.
  5. 如权利要求1-4中任一项所述的化合物，所述阳离子提供基团包含在pH为约4.0或更低的条件下带正电的基团。The compound of any one of claims 1 to 4, wherein the cation providing group comprises a group that is positively charged at a pH of about 4.0 or less.
  6. 如权利要求1-5中任一项所述的化合物，所述阳离子提供基团包含叔胺基团。The compound of any one of claims 1 to 5, wherein the cation providing group comprises a tertiary amine group.
  7. 如权利要求1-6中任一项所述的化合物，所述氢键提供基团包含脲基。The compound according to any one of claims 1 to 6, wherein the hydrogen bond providing group comprises a urea group.
  8. 如权利要求1-7中任一项所述的化合物，所述化合物包含尾部，所述尾部具有疏水性。The compound of any one of claims 1 to 7, comprising a tail, wherein the tail is hydrophobic.
  9. 如权利要求8所述的化合物，所述尾部取代在所述脲基的一个或两个氮原子上。The compound of claim 8, wherein the tail is substituted on one or two nitrogen atoms of the urea group.
  10. 如权利要求8或9所述的化合物，所述尾部包含饱和/或不饱和的脂肪链。The compound according to claim 8 or 9, wherein the tail comprises a saturated and/or unsaturated fatty chain.
  11. 如权利要求8-10中任一项所述的化合物，所述尾部的主链链长为1至20个原子。The compound according to any one of claims 8 to 10, wherein the main chain length of the tail is 1 to 20 atoms.
  12. 如权利要求1-11中任一项所述的化合物，所述化合物包含以下结构(I)，
    The compound according to any one of claims 1 to 11, comprising the following structure (I),
    其中，所述L1为氢键提供基团，所述G1为链长为2至6个原子的连接基团，所述L1任选地被取代，所述G1任选地被取代。Wherein, the L1 is a hydrogen bond providing group, the G1 is a linking group with a chain length of 2 to 6 atoms, the L1 is optionally substituted, and the G1 is optionally substituted.
  13. 如权利要求1-12中任一项所述的化合物，所述化合物包含以下结构(II)，
    The compound according to any one of claims 1 to 12, comprising the following structure (II),
    其中，所述G1为链长为2至6个原子的连接基团，所述G1任选地被取代，所述R1、R2a和R2b各自独立地选自任选取代的取代基。Wherein, the G1 is a linking group with a chain length of 2 to 6 atoms, the G1 is optionally substituted, and the R1 , R2a and R2b are each independently selected from an optionally substituted substituent.
  14. 如权利要求1-13中任一项所述的化合物，所述化合物包含以下结构(IIIa)，
    The compound according to any one of claims 1 to 13, comprising the following structure (IIIa),
    其中，所述G1和G2各自独立地为链长为2至6个原子的连接基团，所述G1和G2各自独立地任选地被取代，所述R1、R2a、R2b、R3、R4a和R4b各自独立地选自任选取代的取代基。wherein G1 and G2 are each independently a linking group having a chain length of 2 to 6 atoms, G1 and G2 are each independently optionally substituted, and R1 , R2a , R2b , R3 , R4a and R4b are each independently selected from optionally substituted substituents.
  15. 如权利要求1-13中任一项所述的化合物，所述化合物包含以下结构(IIIb)，
    The compound according to any one of claims 1 to 13, comprising the following structure (IIIb),
    其中，所述G1为链长为2至6个原子的连接基团，所述G1任选地被取代，所述R1、R2a、R2b、R5和R6各自独立地选自任选取代的取代基。Wherein, the G1 is a linking group with a chain length of 2 to 6 atoms, the G1 is optionally substituted, and the R1 , R2a , R2b , R5 and R6 are each independently selected from optionally substituted substituents.
  16. 如权利要求1-13中任一项所述的化合物，所述化合物包含以下结构(IIIc)，
    The compound according to any one of claims 1 to 13, comprising the following structure (IIIc),
    其中，所述G1和G2各自独立地为链长为2至6个原子的连接基团，所述G1和G2各自独立地任选地被取代，所述R1、R2a、R2b、R7、R8a、R8b和R9各自独立地选自任选取代的取代基。wherein G1 and G2 are each independently a linking group having a chain length of 2 to 6 atoms, G1 and G2 are each independently optionally substituted, and R1 , R2a , R2b , R7 , R8a , R8b and R9 are each independently selected from optionally substituted substituents.
  17. 如权利要求1-13中任一项所述的化合物，所述化合物包含以下结构(IIId)，
    The compound according to any one of claims 1 to 13, comprising the following structure (IIId),
    其中，所述G1、G2、G3和G4各自独立地为链长为2至6个原子的连接基团，所述G1、G2、G3和G4各自独立地任选地被取代，所述H1和H2各自独立地任选地被取代，所述R1、R2a、R2b、R7、R8a、R8b、R10、R11a、R11b、R12、R13a、和R13b各自独立地选自任选取代的取代基。wherein G1 , G2 , G3 and G4 are each independently a linking group having a chain length of 2 to 6 atoms, G1 , G2 , G3 and G4 are each independently optionally substituted, H1 and H2 are each independently optionally substituted, and R1 , R2a , R2b , R7 , R8a , R8b , R10 , R11a , R11b , R12 , R13a , and R13b are each independently selected from optionally substituted substituents.
  18. 如权利要求12-17中任一项所述的化合物，所述G1、G2、G3和G4各自独立地为烃基、脂环基、和/或脂杂环基。The compound according to any one of claims 12 to 17, wherein G 1 , G 2 , G 3 and G 4 are each independently a hydrocarbon group, an alicyclic group, and/or an alicyclic heterocyclic group.
  19. 如权利要求12-18中任一项所述的化合物，所述G1、G2、G3和G4各自独立地为烷基、烯基、和/或炔基。The compound according to any one of claims 12 to 18, wherein G 1 , G 2 , G 3 and G 4 are each independently an alkyl group, an alkenyl group, and/or an alkynyl group.
  20. 如权利要求12-19中任一项所述的化合物，G1、G2、G3和G4中的一个原子各自独立地被N、O或S替换。The compound according to any one of claims 12 to 19, wherein one atom in G 1 , G 2 , G 3 and G 4 is independently replaced by N, O or S.
  21. 如权利要求17-20中任一项所述的化合物，所述H1和H2各自独立地为烃基、脂环基、和/或脂杂环基。The compound according to any one of claims 17 to 20, wherein H1 and H2 are each independently a hydrocarbon group, an alicyclic group, and/or an alicyclic heterocyclic group.
  22. 如权利要求17-21中任一项所述的化合物，所述H1和H2各自独立地为烷基、烯基、和/或炔基。The compound according to any one of claims 17 to 21, wherein H 1 and H 2 are each independently alkyl, alkenyl, and/or alkynyl.
  23. 如权利要求17-22中任一项所述的化合物，H1和H2中的一个原子各自独立地被N、O或S替换。The compound according to any one of claims 17 to 22, wherein one atom of H1 and H2 is independently replaced by N, O or S.
  24. 如权利要求17-23中任一项所述的化合物，所述R1、R2a、R2b、R3、R4a、R4b、R5、R6、R7、R8a、R8b、R9、R10、R11a、R11b、R12、R13a、和R13b各自独立地为烃基、酯基、酰 胺、N’-烷基-N-烯基-脲基、脂环基、脂杂环基、芳基、和/或杂芳基。The compound according to any one of claims 17 to 23, wherein R 1 , R 2a , R 2b , R 3 , R 4a , R 4b , R 5 , R 6 , R 7 , R 8a , R 8b , R 9 , R 10 , R 11a , R 11b , R 12 , R 13a , and R 13b are each independently a hydrocarbon group, an ester group, an acyl group, amine, N'-alkyl-N-alkenyl-urea, alicyclic, alicyclic, aryl, and/or heteroaryl.
  25. 如权利要求17-24中任一项所述的化合物，所述R1、R2a、R2b、R3、R4a、R4b、R5、R6、R7、R8a、R8b、R9、R10、R11a、R11b、R12、R13a、和R13b各自独立地为C1-C20烷基、C2-C40烯基、C2-C45酯基和/或C2-C20炔基。The compound according to any one of claims 17 to 24, wherein R 1 , R 2a , R 2b , R 3 , R 4a , R 4b , R 5 , R 6 , R 7 , R 8a , R 8b , R 9 , R 10 , R 11a , R 11b , R 12 , R 13a , and R 13b are each independently C 1 -C 20 alkyl, C 2 -C 40 alkenyl, C 2 -C 45 ester and/or C 2 -C 20 alkynyl.
  26. 如权利要求17-25中任一项所述的化合物，所述R1、R2a、R2b、R3、R4a、R4b、R5、R6、R7、R8a、R8b、R9、R10、R11a、R11b、R12、R13a、和R13b各自独立地为支化的或非支化的。The compound of any one of claims 17 to 25, wherein R 1 , R 2a , R 2b , R 3 , R 4a , R 4b , R 5 , R 6 , R 7 , R 8a , R 8b , R 9 , R 10 , R 11a , R 11b , R 12 , R 13a , and R 13b are each independently branched or unbranched.
  27. 如权利要求17-26中任一项所述的化合物，所述R1、R2a、R2b、R3、R4a、R4b、R5、R6、R7、R8a、R8b、R9、R10、R11a、R11b、R12、R13a、和R13b各自独立地包含0至4个不饱和键。The compound according to any one of claims 17 to 26, wherein R 1 , R 2a , R 2b , R 3 , R 4a , R 4b , R 5 , R 6 , R 7 , R 8a , R 8b , R 9 , R 10 , R 11a , R 11b , R 12 , R 13a , and R 13b each independently contain 0 to 4 unsaturated bonds.
  28. 如权利要求17-27中任一项所述的化合物，所述R1、R2a、R2b、R3、R4a、R4b、R5、R6、R7、R8a、R8b、R9、R10、R11a、R11b、R12、R13a、和R13b各自独立地被Rx取代，所述Rx为任选取代的取代基。The compound according to any one of claims 17 to 27, wherein R 1 , R 2a , R 2b , R 3 , R 4a , R 4b , R 5 , R 6 , R 7 , R 8a , R 8b , R 9 , R 10 , R 11a , R 11b , R 12 , R 13a , and R 13b are each independently substituted by Rx, and Rx is an optionally substituted substituent.
  29. 如权利要求28所述的化合物，所述Rx为C1-C20烷基、C2-C25酯基、C2-C20烯基、酰胺、脲基、羟基、巯基或氨基。The compound according to claim 28, wherein Rx is a C 1 -C 20 alkyl group, a C 2 -C 25 ester group, a C 2 -C 20 alkenyl group, an amide group, a urea group, a hydroxyl group, a thiol group or an amino group.
  30. 一种化合物，或其药学上可接受的盐，所述化合物选自以下组：



    A compound, or a pharmaceutically acceptable salt thereof, selected from the group consisting of:



  31. 一种组合物，所述组合物包含如权利要求1-30中任一项所述的化合物以及治疗剂。A composition comprising a compound as claimed in any one of claims 1 to 30 and a therapeutic agent.
  32. 如权利要求31所述的组合物，所述组合物还包含选自以下组的物质：中性脂质、类固醇以及聚合物缀合的脂质。The composition of claim 31, further comprising a substance selected from the group consisting of a neutral lipid, a steroid, and a polymer-conjugated lipid.
  33. 如权利要求31或32所述的组合物，所述组合物包含脂质体。The composition of claim 31 or 32, comprising liposomes.
  34. 如权利要求31-33中任一项所述的组合物，所述组合物的直径约为500nm或更小。The composition of any one of claims 31-33, having a diameter of about 500 nm or less.
  35. 如权利要求32-34中任一项所述的组合物，所述类固醇包含胆固醇。The composition of any one of claims 32-34, wherein the steroid comprises cholesterol.
  36. 如权利要求32-35中任一项所述的组合物，所述聚合物缀合的脂质包含聚乙二醇化脂质。The composition of any one of claims 32-35, wherein the polymer-conjugated lipid comprises a PEGylated lipid.
  37. 如权利要求32-36中任一项所述的组合物，所述化合物与所述中性脂质的摩尔比为约2:1至约8:1。The composition of any one of claims 32-36, wherein the molar ratio of the compound to the neutral lipid is from about 2:1 to about 8:1.
  38. 如权利要求32-37中任一项所述的组合物，所述化合物与所述类固醇的摩尔比为约5:1至1:1。The composition of any one of claims 32-37, wherein the molar ratio of the compound to the steroid is about 5:1 to 1:1.
  39. 如权利要求32-38中任一项所述的组合物，所述化合物与所述聚合物缀合的脂质的摩尔比为约100:1至约20:1。The composition of any one of claims 32-38, wherein the molar ratio of the compound to the polymer-conjugated lipid is from about 100:1 to about 20:1.
  40. 如权利要求31-39中任一项所述的组合物，所述治疗剂包含核酸。The composition of any one of claims 31-39, wherein the therapeutic agent comprises a nucleic acid.
  41. 如权利要求31-40中任一项所述的组合物，所述治疗剂包含CRISPR系统的组分和/或编码CRISPR系统的组分的核酸。The composition of any one of claims 31-40, wherein the therapeutic agent comprises a component of a CRISPR system and/or a nucleic acid encoding a component of a CRISPR system.
  42. 如权利要求31-41中任一项所述的组合物，所述治疗剂包含guide RNA和/或编码Cas酶的核酸。A composition as described in any one of claims 31-41, wherein the therapeutic agent comprises a guide RNA and/or a nucleic acid encoding a Cas enzyme.
  43. 一种细胞，所述细胞包含如权利要求1-30中任一项所述的化合物和/或如权利要求31-42中任一项所述的组合物。A cell comprising the compound according to any one of claims 1 to 30 and/or the composition according to any one of claims 31 to 42.
  44. 一种试剂盒，所述试剂盒包含如权利要求1-30中任一项所述的化合物、如权利要求31-42中任一项所述的组合物和/或如权利要求43所述的细胞。A kit comprising the compound according to any one of claims 1 to 30, the composition according to any one of claims 31 to 42, and/or the cell according to claim 43.
  45. 一种施用治疗剂的方法，所述方法包含提供权利要求1-30中任一项所述的化合物、如权利要求31-42中任一项所述的组合物、如权利要求43所述的细胞和/或如权利要求44所述的试剂盒，所述治疗剂与所述化合物混合，或所述治疗剂存在于所述组合物、所述细胞和/或所述试剂盒中。A method of administering a therapeutic agent, the method comprising providing a compound as described in any one of claims 1-30, a composition as described in any one of claims 31-42, a cell as described in claim 43, and/or a kit as described in claim 44, wherein the therapeutic agent is mixed with the compound or the therapeutic agent is present in the composition, the cell, and/or the kit.
  46. 一种递送核酸的方法，所述方法包含提供权利要求1-30中任一项所述的化合物、如权利要求31-42中任一项所述的组合物、如权利要求43所述的细胞和/或如权利要求44所述 的试剂盒，所述核酸与所述化合物混合，或所述核酸存在于所述组合物、所述细胞和/或所述试剂盒中。A method for delivering a nucleic acid, the method comprising providing a compound according to any one of claims 1 to 30, a composition according to any one of claims 31 to 42, a cell according to claim 43, and/or a cell according to claim 44. The nucleic acid is mixed with the compound, or the nucleic acid is present in the composition, the cell and/or the kit.
  47. 一种治疗和/或预防疾病或病症的方法，所述方法包含提供权利要求1-30中任一项所述的化合物、如权利要求31-42中任一项所述的组合物、如权利要求43所述的细胞和/或如权利要求44所述的试剂盒。 A method for treating and/or preventing a disease or condition, the method comprising providing a compound according to any one of claims 1 to 30, a composition according to any one of claims 31 to 42, a cell according to claim 43 and/or a kit according to claim 44.
PCT/CN2023/129240 2022-11-02 2023-11-02 Compound for delivery system and use thereof WO2024094098A1 (en)

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