WO2024032753A1 - Nitrogen-containing chain compound, preparation method therefor, composition containing said compound, and use thereof - Google Patents

Nitrogen-containing chain compound, preparation method therefor, composition containing said compound, and use thereof Download PDF

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WO2024032753A1
WO2024032753A1 PCT/CN2023/112482 CN2023112482W WO2024032753A1 WO 2024032753 A1 WO2024032753 A1 WO 2024032753A1 CN 2023112482 W CN2023112482 W CN 2023112482W WO 2024032753 A1 WO2024032753 A1 WO 2024032753A1
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alkyl group
group
butyl
substituted
unsubstituted
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PCT/CN2023/112482
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French (fr)
Chinese (zh)
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林金钟
卢静
俞航
姜婷
张凡
张博阳
郭俊香
陈林
王冰
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上海蓝鹊生物医药有限公司
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    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K31/00—Medicinal preparations containing organic active ingredients
    • A61K31/70—Carbohydrates; Sugars; Derivatives thereof
    • A61K31/7088—Compounds having three or more nucleosides or nucleotides
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • 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
    • 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
    • A61K47/183—Amino acids, e.g. glycine, EDTA or aspartame
    • 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
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K9/00—Medicinal preparations characterised by special physical form
    • A61K9/48—Preparations in capsules, e.g. of gelatin, of chocolate
    • A61K9/50—Microcapsules having a gas, liquid or semi-solid filling; Solid microparticles or pellets surrounded by a distinct coating layer, e.g. coated microspheres, coated drug crystals
    • A61K9/51—Nanocapsules; Nanoparticles
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K9/00—Medicinal preparations characterised by special physical form
    • A61K9/48—Preparations in capsules, e.g. of gelatin, of chocolate
    • A61K9/50—Microcapsules having a gas, liquid or semi-solid filling; Solid microparticles or pellets surrounded by a distinct coating layer, e.g. coated microspheres, coated drug crystals
    • A61K9/51—Nanocapsules; Nanoparticles
    • A61K9/5107—Excipients; Inactive ingredients
    • A61K9/5123—Organic compounds, e.g. fats, sugars
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07C—ACYCLIC OR CARBOCYCLIC COMPOUNDS
    • C07C229/00—Compounds containing amino and carboxyl groups bound to the same carbon skeleton
    • C07C229/02—Compounds containing amino and carboxyl groups bound to the same carbon skeleton having amino and carboxyl groups bound to acyclic carbon atoms of the same carbon skeleton
    • C07C229/04—Compounds containing amino and carboxyl groups bound to the same carbon skeleton having amino and carboxyl groups bound to acyclic carbon atoms of the same carbon skeleton the carbon skeleton being acyclic and saturated
    • C07C229/06—Compounds containing amino and carboxyl groups bound to the same carbon skeleton having amino and carboxyl groups bound to acyclic carbon atoms of the same carbon skeleton the carbon skeleton being acyclic and saturated having only one amino and one carboxyl group bound to the carbon skeleton
    • C07C229/10—Compounds containing amino and carboxyl groups bound to the same carbon skeleton having amino and carboxyl groups bound to acyclic carbon atoms of the same carbon skeleton the carbon skeleton being acyclic and saturated having only one amino and one carboxyl group bound to the carbon skeleton the nitrogen atom of the amino group being further bound to acyclic carbon atoms or to carbon atoms of rings other than six-membered aromatic rings
    • C07C229/16—Compounds containing amino and carboxyl groups bound to the same carbon skeleton having amino and carboxyl groups bound to acyclic carbon atoms of the same carbon skeleton the carbon skeleton being acyclic and saturated having only one amino and one carboxyl group bound to the carbon skeleton the nitrogen atom of the amino group being further bound to acyclic carbon atoms or to carbon atoms of rings other than six-membered aromatic rings to carbon atoms of hydrocarbon radicals substituted by amino or carboxyl groups, e.g. ethylenediamine-tetra-acetic acid, iminodiacetic acids
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07C—ACYCLIC OR CARBOCYCLIC COMPOUNDS
    • C07C2601/00—Systems containing only non-condensed rings
    • C07C2601/04—Systems containing only non-condensed rings with a four-membered ring
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07C—ACYCLIC OR CARBOCYCLIC COMPOUNDS
    • C07C2601/00—Systems containing only non-condensed rings
    • C07C2601/06—Systems containing only non-condensed rings with a five-membered ring
    • C07C2601/08—Systems containing only non-condensed rings with a five-membered ring the ring being saturated
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07C—ACYCLIC OR CARBOCYCLIC COMPOUNDS
    • C07C2601/00—Systems containing only non-condensed rings
    • C07C2601/12—Systems containing only non-condensed rings with a six-membered ring
    • C07C2601/14—The ring being saturated
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07C—ACYCLIC OR CARBOCYCLIC COMPOUNDS
    • C07C2601/00—Systems containing only non-condensed rings
    • C07C2601/18—Systems containing only non-condensed rings with a ring being at least seven-membered
    • C07C2601/20—Systems containing only non-condensed rings with a ring being at least seven-membered the ring being twelve-membered

Definitions

  • the present invention relates to nitrogen-containing chain compounds, methods for their preparation, compositions containing them and their uses.
  • Nucleic acid drugs are an important direction in current basic and applied research. Nucleic acid drugs can be used for the prevention and/or treatment of viral and bacterial infectious diseases, tumors, metabolic diseases, etc. Their production costs are lower and the cycle is shorter, which is conducive to the rapid development of personalized drugs. However, nucleic acids are negatively charged macromolecules that are difficult to penetrate through cell membranes. At the same time, nucleic acids have poor stability. By developing various nucleic acid packaging and delivery systems, the instability of nucleic acid drugs can be overcome to a certain extent and their delivery efficiency can be improved.
  • Lipid nanoparticles have been shown to be useful as vehicles for delivering bioactive substances, such as small molecule drugs, proteins, and nucleic acids, into cells and/or intracellular compartments.
  • Optimizing the nucleic acid drug delivery system by designing and optimizing the types and amounts of each component in lipid nanoparticles is of great significance to improve the efficacy of nucleic acid drug prevention and treatment, especially for the delivery of RNA preventive and/or therapeutic agents.
  • Lipid compounds and related methods and compositions are of great significance to improve the efficacy of nucleic acid drug prevention and treatment, especially for the delivery of RNA preventive and/or therapeutic agents.
  • the present invention aims to provide a new ionizable lipid compound that can be used to deliver nucleic acid drugs, and to increase the types of ionizable lipid compounds and the selection of nucleic acid preventive and/or therapeutic agent delivery carriers.
  • the present invention provides a nitrogen-containing chain compound, its preparation method, compositions containing the same and applications.
  • the composition of the present invention can be used to efficiently deliver nucleic acid drugs.
  • the present invention provides a nitrogen-containing chain compound shown in formula I or a pharmaceutically acceptable salt thereof,
  • X is a C 1 -C 6 alkylene group or a C 3 -C 6 cycloalkylene group
  • Y is an unsubstituted or C 1 -C 10 alkylene group substituted by 1, 2 or 3 Y 1-1 ;
  • Each Y 1-1 is independently hydroxyl, halogen or C 1 -C 6 alkyl
  • Z is an unsubstituted or C 1 -C 10 alkylene group substituted by 1, 2 or 3 Z 1-1 ;
  • Each Z 1-1 is independently hydroxyl, halogen or C 1 -C 6 alkyl
  • a 1 and A 2 are independently
  • R 1 is a C 1 -C 20 alkyl group that is unsubstituted or substituted by 1, 2 or 3 R 1-1 or unsubstituted or substituted by 1, 2 or 3 R 1-2 C 2 -C 20 alkenyl;
  • Each R 1-1 is independently a hydroxyl group, a C 1 -C 10 alkyl group, a C 3 -C 15 cycloalkyl group that is unsubstituted or substituted by 1, 2 or 3 R 1-1-4 ,
  • R 1-1-1 is a C 1 -C 6 alkyl group
  • R 1-1-2 is a C 1 -C 6 alkyl group
  • R 1-1-3 is a C 1 -C 6 alkyl group
  • each R 1-1-4 is independently a C 1 -C 6 alkyl group
  • Each R 1-2 is independently a hydroxyl group or a C 1 -C 10 alkyl group
  • R 2 is a C 1 -C 20 alkyl group that is unsubstituted or substituted by 1, 2 or 3 R 2-1 or unsubstituted or substituted by 1, 2 or 3 R 2-2 C 2 -C 20 alkenyl;
  • Each R 2-1 is independently a hydroxyl group, a C 1 -C 10 alkyl group, a C 3 -C 15 cycloalkyl group that is unsubstituted or substituted by 1, 2 or 3 R 2-1-4 ,
  • R 2-1-1 is a C 1 -C 6 alkyl group
  • R 2-1-2 is a C 1 -C 6 alkyl group
  • R 2-1-3 is a C 1 -C 6 alkyl group
  • each R 2-1-4 is independently a C 1 -C 6 alkyl group
  • Each R 2-2 is independently a hydroxyl group or a C 1 -C 10 alkyl group
  • R 1 is a C 1 -C 20 alkyl group substituted by 1, 2 or 3 hydroxyl groups, unsubstituted, or Cyclopentane substituted with R 1-1-4 , cyclopentane unsubstituted or substituted by 1, 2 or 3 R 1-1-4 ,
  • the C 1 -C 6 alkylene group may be a C 1 -C 4 alkylene group, preferably For example
  • the C 3 -C 6 cycloalkylene group can be
  • the C 3 -C 6 cycloalkylene group can be
  • the C 1 -C 10 alkylene group may be a C 4 -C 6 alkylene group, preferably For example
  • the halogen in each Y 1-1 , can be fluorine, chlorine, bromine or iodine.
  • the C 1 -C 6 alkylene group in each Y 1-1 , can be
  • the C 1 -C 10 alkylene group may be a C 5 -C 8 alkylene group, preferably For example
  • the halogen may be fluorine, chlorine, bromine or iodine.
  • the C 1 -C 6 alkylene group in each Z 1-1 , can be
  • the C 1 -C 20 alkyl group can be a C 1 -C 15 alkyl group, preferably a straight chain alkyl group, such as ethyl, propyl, hexyl, octyl, Nonyl or dodecyl.
  • the C 2 -C 20 alkenyl group may be a C 6 -C 18 alkenyl group, preferably a linear alkenyl group; the C 2 -C 20 alkenyl group may contain 1 to 4 double bonds.
  • the C 1 -C 10 alkyl group in each R 1-1 , can be a C 1 -C 4 alkyl group, preferably methyl, ethyl, n-propyl, isopropyl, n-propyl, etc. Butyl, tert-butyl, isobutyl or sec-butyl, for example methyl, ethyl, isopropyl, tert-butyl or n-butyl.
  • the C 3 -C 15 cycloalkyl group can be a C 3 -C 12 cycloalkyl group, preferably a C 4 -C 8 cycloalkyl group or a cycloalkyl group. group, such as cyclopentyl, cyclohexyl or cycloalkyl.
  • the C 1 -C 6 alkyl group can be a C 1 -C 4 alkyl group, preferably methyl, ethyl, n-propyl, isopropyl, n-butyl, tert-butyl, iso-butyl or sec-butyl.
  • the C 1 -C 6 alkyl group can be a C 1 -C 4 alkyl group, preferably methyl, ethyl, n-propyl, isopropyl, n-butyl, tert-butyl, iso-butyl or sec-butyl.
  • the C 1 -C 6 alkyl group can be a C 1 -C 4 alkyl group, preferably methyl, ethyl, n-propyl, isopropyl, n-butyl, tert-butyl, iso-butyl or sec-butyl.
  • the C 1 -C 6 alkyl group can be a C 1 -C 4 alkyl group, preferably methyl, ethyl, n-propyl, isopropyl , n-butyl, tert-butyl, isobutyl or sec-butyl.
  • the C 1 -C 10 alkyl group can be a C 1 -C 6 alkyl group, preferably methyl, ethyl, n-propyl, Isopropyl, n-butyl, tert-butyl, isobutyl or sec-butyl.
  • the C 1 -C 20 alkyl group can be a C 1 -C 15 alkyl group, preferably a straight chain alkyl group, such as ethyl, propyl, hexyl, octyl, Nonyl or dodecyl.
  • the C 2 -C 20 alkenyl group may be a C 6 -C 18 alkenyl group, preferably a linear alkenyl group; the C 2 -C 20 alkenyl group may contain 1 to 4 double bonds.
  • the C 1 -C 10 alkyl group can be a C 1 -C 8 alkyl group, preferably methyl, ethyl, n-propyl, isopropyl, n-propyl, etc.
  • the C 3 -C 15 cycloalkyl group can be a C 3 -C 12 cycloalkyl group, preferably a C 4 -C 8 cycloalkyl group or a cycloalkyl group. group, such as cyclopentyl, cyclohexyl or cycloalkyl.
  • the C 1 -C 6 alkyl group can be a C 1 -C 4 alkyl group, preferably methyl, ethyl, n-propyl, isopropyl, n-butyl, tert-butyl, iso-butyl or sec-butyl.
  • the C 1 -C 6 alkyl group can be a C 1 -C 4 alkyl group, preferably methyl, ethyl, n-propyl, isopropyl, n-butyl, tert-butyl, iso-butyl or sec-butyl.
  • the C 1 -C 6 alkyl group can be a C 1 -C 4 alkyl group, preferably methyl, ethyl, n-propyl, isopropyl, n-butyl, tert-butyl, iso-butyl or sec-butyl.
  • the C 1 -C 6 alkyl group can be a C 1 -C 4 alkyl group, preferably methyl, ethyl, n-propyl, isopropyl , n-butyl, tert-butyl, isobutyl or sec-butyl.
  • the C 1 -C 10 alkyl group can be a C 1 -C 6 alkyl group, preferably methyl, ethyl, n-propyl, isopropyl, n-propyl, etc. Butyl, tert-butyl, isobutyl or sec-butyl.
  • the nitrogen-containing chain compound represented by formula I is a compound represented by formula Ia, Ib, Ic, Id or Ie.
  • a 1 is where a is connected to R 1 and b is connected to Y, or a is connected to Y and b is connected to R 1 .
  • a 2 is Where a is connected to R 2 and b is connected to Z, or a is connected to Z and b is connected to R 2 .
  • R 1 and R 2 are the same.
  • X is a C 1 -C 4 alkylene group or a C 3 -C 6 cycloalkylene group
  • Y is a C 1 -C 10 alkylene group
  • Z is a C 1 -C 10 alkylene group
  • a 1 and A 2 are independently
  • R 1 is a C 1 -C 20 alkyl group that is unsubstituted or substituted by 1, 2 or 3 R 1-1 ;
  • Each R 1-1 is independently a hydroxyl group, a C 1 -C 10 alkyl group or a C 3 -C 15 cycloalkyl group that is unsubstituted or substituted by 1, 2 or 3 R 1-1-4 ;
  • Each R 1-1-4 is independently a C 1 -C 6 alkyl group
  • R 2 is a C 1 -C 20 alkyl group that is unsubstituted or substituted by 1, 2 or 3 R 2-1 ;
  • Each R 2-1 is independently a hydroxyl group, a C 1 -C 10 alkyl group or a C 3 -C 15 cycloalkyl group that is unsubstituted or substituted by 1, 2 or 3 R 2-1-4 ;
  • Each R 2-1-4 is independently a C 1 -C 6 alkyl group.
  • X is a C 1 -C 4 alkylene group or a C 3 -C 6 cycloalkylene group
  • Y is a C 4 -C 8 alkylene group
  • Z is a C 4 -C 8 alkylene group
  • a 1 is Where a is connected to R 1 , b is connected to Y;
  • a 2 is Among them, a is connected to R 2 , and b is connected to Z;
  • R 1 is a C 1 -C 20 alkyl group that is unsubstituted or substituted by 1, 2 or 3 R 1-1 ;
  • Each R 1-1 is independently a hydroxyl group, a C 1 -C 6 alkyl group or a C 3 -C 6 cycloalkyl group that is unsubstituted or substituted by 1, 2 or 3 R 1-1-4 ;
  • Each R 1-1-4 is independently a C 1 -C 6 alkyl group
  • R 2 is a C 1 -C 20 alkyl group that is unsubstituted or substituted by 1, 2 or 3 R 2-1 ;
  • Each R 2-1 is independently a hydroxyl group, a C 1 -C 6 alkyl group or a C 3 -C 6 cycloalkyl group that is unsubstituted or substituted by 1, 2 or 3 R 2-1-4 ;
  • Each R 2-1-4 is independently a C 1 -C 6 alkyl group
  • R 1 is a C 1 -C 20 alkyl group substituted by 1, 2 or 3 hydroxyl groups, unsubstituted, or R 1-1-4 substituted cyclopentane
  • X is a C 1 -C 4 alkylene group
  • Y is a C 4 -C 8 alkylene group
  • Z is a C 4 -C 8 alkylene group
  • a 1 is Where a is connected to R 1 , b is connected to Y;
  • a 2 is Among them, a is connected to R 2 , and b is connected to Z;
  • R 1 is a C 1 -C 15 alkyl group substituted by 1, 2 or 3 hydroxyl groups or cyclopentane
  • R 2 is a C 1 -C 15 alkyl group substituted by 1, 2 or 3 R 2-1 ;
  • Each R 2-1 is independently a C 1 -C 10 alkyl group.
  • X is a C 1 -C 4 alkylene group or a C 3 -C 6 cycloalkylene group
  • Y is a C 4 -C 8 alkylene group
  • Z is a C 4 -C 8 alkylene group
  • a 1 is Where a is connected to R 1 , b is connected to Y;
  • a 2 is Among them, a is connected to R 2 , and b is connected to Z;
  • R 1 is a C 1 -C 20 alkyl group that is unsubstituted or substituted by 1, 2 or 3 R 1-1 ;
  • Each R 1-1 is independently a hydroxyl group, a C 1 -C 6 alkyl group or a C 3 -C 6 cycloalkyl group that is unsubstituted or substituted by 1, 2 or 3 R 1-1-4 ;
  • Each R 1-1-4 is independently a C 1 -C 6 alkyl group
  • R 2 is a C 1 -C 20 alkyl group that is unsubstituted or substituted by 1, 2 or 3 R 2-1 ;
  • Each R 2-1 is independently a hydroxyl group, a C 1 -C 8 alkyl group or a C 3 -C 6 cycloalkyl group that is unsubstituted or substituted by 1, 2 or 3 R 2-1-4 ;
  • Each R 2-1-4 is independently a C 1 -C 6 alkyl group
  • R 1 is a C 1 -C 20 alkyl group substituted by 1, 2 or 3 hydroxyl groups, unsubstituted, or R 1-1-4 substituted cyclopentane
  • X is a C 1 -C 4 alkylene group
  • Y is a C 4 -C 8 alkylene group
  • Z is a C 4 -C 8 alkylene group
  • a 1 is Where a is connected to R 1 , b is connected to Y;
  • a 2 is Among them, a is connected to R 2 , and b is connected to Z;
  • R 1 is a C 1 -C 15 alkyl group substituted by 1, 2 or 3 hydroxyl groups or cyclopentane
  • R 2 is a C 1 -C 15 alkyl group substituted by 1, 2 or 3 R 2-1 ;
  • Each R 2-1 is independently a C 1 -C 10 alkyl group or a C 3 -C 6 cycloalkyl group that is unsubstituted or substituted by 1, 2 or 3 R 1-1-4 ;
  • Each R 2-1-4 is independently a C 1 -C 6 alkyl group.
  • X is a C 1 -C 4 alkylene group or a C 3 -C 6 cycloalkylene group.
  • Y is a C 1 -C 10 alkylene group.
  • Z is a C 1 -C 10 alkylene group.
  • R 1 is a C 1 -C 20 alkyl group that is unsubstituted or substituted by 1, 2 or 3 R 1-1 .
  • each R 1-1 is independently a hydroxyl group, a C 1 -C 10 alkyl group or an unsubstituted C 3 -C substituted by 1, 2 or 3 R 1-1-4 15 cycloalkyl.
  • R 2 is a C 1 -C 20 alkyl group that is unsubstituted or substituted by 1, 2 or 3 R 2-1 .
  • each R 2-1 is independently a hydroxyl group, a C 1 -C 10 alkyl group or an unsubstituted C 3 -C substituted by 1, 2 or 3 R 2-1-4 15 cycloalkyl.
  • Y is a C 4 -C 8 alkylene group.
  • Z is a C 4 -C 8 alkylene group.
  • a 1 is Where a is connected to R 1 and b is connected to Y.
  • a 2 is Where a is connected to R 2 and b is connected to Z.
  • each R 1-1 is independently a hydroxyl group, a C 1 -C 6 alkyl group or an unsubstituted C 3 -C substituted by 1, 2 or 3 R 1-1-4 6 cycloalkyl.
  • each R 2-1 is independently a hydroxyl group, a C 1 -C 6 alkyl group or an unsubstituted C 3 -C substituted by 1, 2 or 3 R 2-1-4 6 cycloalkyl.
  • R 1 is a C 1 -C 20 alkyl group substituted by 1, 2 or 3 hydroxyl groups, unsubstituted or One, two or three R 1-1-4 substituted cyclopentane,
  • R 1 is
  • R 2 is
  • the nitrogen-containing chain compound shown in Formula I is any of the following compounds:
  • the invention also provides a method for preparing a nitrogen-containing chain compound represented by formula I, which includes the following steps: in a solvent, in the presence of a base and an iodide salt, a compound represented by formula I-1 is mixed with a compound represented by formula I
  • the compound represented by -2 can undergo a coupling reaction as shown in the following formula;
  • M is halogen, and X, Y, Z, A 1 , A 2 , R 1 and R 2 are as described above.
  • the halogen may be fluorine, chlorine, bromine or iodine, such as bromine.
  • the molar ratio of the compound represented by Formula I-1 to the compound represented by Formula I-2 may be 1:(1-2), Example 1:1.2.
  • the base is a conventional base in this field.
  • the base may be a basic carbonate (the cation in the salt is an alkali metal ion and the anion is carbonate), such as K 2 CO 3 .
  • the molar ratio of the compound represented by Formula I-1 to the base may be 1:(1-5), such as 1:3.5 or 1:2.
  • the iodized salt is a conventional iodized salt in this field.
  • the iodized salt may be a basic iodized salt, such as KI.
  • the molar ratio of the compound represented by Formula I-1 to the iodide salt may be 1:(1-2), such as 1:1.2 or 1:1.
  • the solvent is a conventional solvent in this field, and the solvent can be an ether solvent or/and a nitrile solvent.
  • the ether solvent may be methyl tert-butyl ether.
  • the nitrile solvent may be acetonitrile.
  • the volume ratio of the nitrile solvent to the ether solvent can be 1:1.
  • the mass-to-volume ratio of the compound represented by Formula I-1 to the solvent may be 10 mg/mL-65 mg/mL, such as 15 mg/mL, 25 mg/mL, 40 mg/mL, 58.5 mg/mL, 20mg/L, 22mg/L or 34mg/L.
  • the reaction temperature of the coupling reaction is a conventional reaction temperature in this field, preferably 70°C to 90°C, such as 80°C.
  • the present invention also provides a lipid carrier, which includes substance Z, which is a compound represented by Formula I as described above or a pharmaceutically acceptable salt thereof.
  • the lipid carrier further includes a diluent.
  • the diluent may be phosphate buffer or Tris buffer, etc.
  • the lipid carrier further includes phospholipids.
  • the phospholipid can be a conventional phospholipid in the art, which is an amphoteric auxiliary molecule that contributes to the fusion of lipid particles and cell membranes.
  • the phospholipid may be a phospholipid molecule with a charged polar end and a non-polar end of the fatty chain, such as distearoyl phosphatidylcholine (DSPC), dimyristoyl phosphocholine (DMPC), dioleoyl phosphocholine alkali (DOPC), palmitoyl phosphocholine (DPPC), 1,2-distearoyl phosphocholine (DSPC), undecanoyl phosphocholine (DUPC) or palmitoyl phosphocholine (POPC), etc.
  • DSPC distearoyl phosphatidylcholine
  • DMPC dimyristoyl phosphocholine
  • DOPC dioleoyl phosphocholine alkali
  • DPPC palmitoyl phosphocholine
  • DUPC unde
  • the lipid carrier also includes PEG lipid (polyethylene glycol modified lipid).
  • the PEG lipid may be a lipid molecule modified with a polyethylene glycol hydrophilic end.
  • the PEG lipid is preferably selected from the group consisting of PEG-modified phosphatidylethanolamine, PEG-modified phosphatidic acid, PEG-modified ceramide, PEG-modified dialkylamine, PEG-modified diacylglycerol and PEG-modified dialkylglycerol.
  • PEG-modified dimyristoylglycerol DMG-PEG2000
  • the lipid carrier further includes sterols.
  • the sterols can be conventional sterols in this field, and the sterols include animal, plant or fungal sterols.
  • the sterol is selected from one or more of cholesterol, sitosterol, ergosterol, campesterol, stigmasterol, brassinosterol, tomatine, ursolic acid and ⁇ -tocopherol, such as cholesterol and the like.
  • the molar ratio of the substance Z to the sterol is 0.5-5:1, preferably 0.5-3:1, for example, 0.6-2:1.
  • the molar ratio of the substance Z to the phospholipid is 1 to 15:1, preferably 2 to 8:1, such as 3 to 6:1.
  • the molar ratio of the substance Z to the PEG lipid is 20 to 130:1, preferably 20 to 80:1, such as 20 to 40:1.
  • the molar content of substance Z is about 30 mol% to 60 mol%.
  • the molar content means the percentage of a certain substance in the total mass of the lipid carrier, and the sum of the molar contents of each component in the lipid carrier does not exceed 100 mol%. In a certain preferred embodiment, the molar content of the phospholipid is about 0 mol% to 30 mol%.
  • the molar content of the sterol is about 15 mol% to 55 mol%.
  • the molar content of the PEG lipid is about 0 mol% to 10 mol%.
  • the lipid carrier consists of the substance Z, the diluent, the phospholipid, the PEG lipid and the sterol.
  • the present invention also provides a lipid nanoparticle, which includes a therapeutic agent and/or a preventive agent and the aforementioned lipid carrier.
  • the therapeutic agent and/or preventive agent may be one or two or more nucleic acids.
  • the nucleic acid may be a conventional nucleic acid in the art.
  • the therapeutic and/or preventive agent may be single-stranded deoxyribonucleic acid (DNA), double-stranded DNA, small interfering RNA (siRNA), asymmetric double-stranded small interfering RNA (aiRNA), microRNA (miRNA), small RNA Clamp RNA (shRNA), circular RNA (circRNA), transfer RNA (tRNA), messenger RNA (mRNA) and other forms of nucleic acid molecules known in the art, preferably mRNA, such as firefly luciferase (Fluc) mRNA or SARS-CoV-2 spike protein (Spike) mRNA.
  • mRNA such as firefly luciferase (Fluc) mRNA or SARS-CoV-2 spike protein (Spike) mRNA.
  • the nitrogen to phosphorus ratio in the lipid nanoparticles can be 2:1 to 30:1, and the nitrogen to phosphorus ratio of the composition refers to the ionizable nitrogen in one or more ionizable lipid compounds.
  • the ratio of the number of moles of atoms to the number of moles of phosphate groups in RNA Preferably, it is 2:1 to 20:1, such as 3:1 to 20:1, and another example is 3:1 to 16:1.
  • the mass ratio of the lipid carrier to the therapeutic agent and/or preventive agent can be 3 to 80:1, preferably 6 to 60:1.
  • the particle size (average particle size) of the lipid nanoparticles can be 10 to 250 nm, for example, 40 to 250 nm, further 50 to 250 nm, or 40 to 150 nm, or 60 to 150 nm.
  • the lipid carrier encapsulates the therapeutic agent and/or preventive agent.
  • the present invention also provides a composition, which includes substance Z, which is a compound represented by Formula I as described above or a pharmaceutically acceptable salt thereof.
  • the composition further includes one or more of diluent, phospholipid, PEG lipid, sterol and therapeutic and/or preventive agent.
  • the diluent, phospholipid, PEG lipid, sterol and therapeutic agent and/or preventive agent are as described above.
  • the substance Z and one or more of the diluent, phospholipid, PEG lipid and sterol form a lipid carrier as described above.
  • the lipid carrier and the therapeutic agent and/or preventive agent form lipid nanoparticles as described above.
  • the encapsulation rate of the therapeutic agent and/or preventive agent in the composition is at least 50%, preferably at least 70%.
  • the polydispersity index of the composition is not higher than 0.5, for example, not higher than 0.3.
  • halogen refers to fluorine, chlorine, bromine or iodine.
  • pharmaceutically acceptable means relatively nontoxic, safe, and suitable for use by patients.
  • pharmaceutically acceptable salt refers to a salt obtained by reacting a compound with a pharmaceutically acceptable acid or base.
  • base addition salts can be obtained by contacting the compound with a sufficient amount of a pharmaceutically acceptable base in a suitable inert solvent.
  • Pharmaceutically acceptable base addition salts include, but are not limited to, sodium salts, potassium salts, calcium salts, aluminum salts, magnesium salts, bismuth salts, ammonium salts, etc.
  • acid addition salts can be obtained by contacting the compound with a sufficient amount of a pharmaceutically acceptable acid in a suitable inert solvent.
  • Pharmaceutically acceptable acid addition salts include, but are not limited to: hydrochloride, sulfate, methanesulfonate, etc.
  • hydrochloride sulfate, methanesulfonate, etc.
  • the "-" at the end of a group means that the group is connected to the rest of the molecule through that site.
  • alkyl refers to a linear or branched, saturated monovalent hydrocarbon group having a specified number of carbon atoms (eg, C 1 to C 6 ).
  • Alkyl groups include, but are not limited to: methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, sec-butyl, tert-butyl, n-pentyl, n-hexyl, etc.
  • alkylene is a divalent group connected to the rest of the molecule by two single bonds and is otherwise defined in the same manner as the term "alkyl”.
  • cycloalkyl refers to a saturated monocyclic cyclic group having a specified number of carbon atoms (eg, C 3 to C 6 ) and consisting only of carbon atoms. Cycloalkyl groups include, but are not limited to, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, and the like.
  • alkenyl refers to a straight or branched chain hydrocarbon group containing one or more carbon-carbon double bonds (eg, 1, 2, or 3 carbon-carbon double bonds).
  • any variable (such as the group R 1-1 ) appears multiple times in the definition of a compound, their definitions are independent of each other and do not affect each other.
  • a C 6 to C 10 aryl group substituted by 3 R 1-1 means that the C 6 to C 10 aryl group will be substituted by 3 R 1-1 .
  • the definitions of the three R 1-1 are independent of each other. No impact.
  • the reagents and raw materials used in the present invention are all commercially available.
  • the positive and progressive effect of the present invention is that the present invention provides a nitrogen-containing chain compound shown in Formula I, which has a novel structure and can be used to prepare lipid nanoparticles.
  • Lipid nanoparticles containing nitrogen-containing chain compounds as shown in Formula I have a low polydispersity index and can transport mRNA efficiently.
  • Figure 1 shows the gel electrophoresis pattern of each Fluc-mRNA LNP prepared in Implementation 23.
  • Figure 2 shows the chemiluminescence intensity of 293FT cells in Example 25 after co-culture with each Fluc-mRNA LNP for 18-24 hours.
  • Figure 3 shows the Spike protein expression of 293FT cells and each Spike-mRNA LNP after co-culture for 24 hours in Example 25, and the PBS group was used as a negative control.
  • 8-Bromooctanoic acid purchased from Jiangsu Aikang, purity: 98%;
  • Heptadecan-9-ol purchased from Beijing Fuyuzhou Technology Co., Ltd., purity: 98%;
  • DCC dicyclohexylcarbodiimide, purchased from Adamas Reagent Co., Ltd., product number 012041444, purity: RG, 99%;
  • DMAP 4-dimethylaminopyridine, purchased from Adamas Reagent Co., Ltd., product number: 01271081, purity: RG, 99%;
  • DCM dichloromethane, purchased from Shanghai Titan Technology Co., Ltd., item number: 01111853, purity: AR, ⁇ 99.5%;
  • Ethanolamine purchased from Shanghai Yien Chemical Technology Co., Ltd., product number: R016710, purity: AR, 99%;
  • Acetonitrile purchased from Shanghai Titan Technology Co., Ltd., product number: 01111797, purity: AR, ⁇ 99.0%;
  • 6-Bromohexanoic acid purchased from Adamas Reagent Co., Ltd., item number: 01073739, purity: RG, 98%+;
  • K 2 CO 3 purchased from Shanghai Yien Chemical Technology Co., Ltd., item number: RH425011, purity: AR, 99%;
  • Methyl tert-butyl ether purchased from Shanghai Titan Technology Co., Ltd., product number: 01030342, purity: AR, ⁇ 99.0%;
  • Isodecanol purchased from Shanghai Aladdin Biochemical Technology Co., Ltd., product number: I298943, purity: 98%;
  • Diatomite purchased from Shanghai Titan Technology Co., Ltd., item number: 01589000, purity: premium grade, ⁇ 89.0%, 200 mesh;
  • Ethyl acetate purchased from Shanghai Titan Technology Co., Ltd., product number: 01153552, purity: AR, ⁇ 99.5%;
  • 3,5,5-Trimethyl-1-hexanol purchased from Shanghai Aladdin Biochemical Technology Co., Ltd., product number: T162750, purity: GC, >85.0%;
  • 2-Ethylhexanol purchased from Shanghai Yien Chemical Technology Co., Ltd., product number: R016621, purity: AR, 99%;
  • Anhydrous sodium sulfate purchased from Shanghai Titan Technology Co., Ltd., product number: 01224581, purity: AR, ⁇ 99.0%;
  • Ferric chloride purchased from Shanghai McLean Biochemical Technology Co., Ltd., item number: I811935, purity: AR, 99%;
  • Pyridine purchased from Shanghai McLean Biochemical Technology Co., Ltd., product number: P816288, purity: AR, 99%;
  • 1,2-Epoxydodecane purchased from Shanghai McLean Biochemical Technology Co., Ltd., product number: E808874, purity: 95%;
  • Cyclohexylmethanol purchased from Shanghai Yien Chemical Technology Co., Ltd., product number: R014045, purity: 99%;
  • Isononyl alcohol purchased from Guangdong Wengjiang Chemical Reagent Co., Ltd.;
  • 8-Methyl-1-decanol purchased from Nanjing Yilan Biotechnology Co., Ltd., purity: 97%;
  • reaction solution was filtered through diatomaceous earth and spun to dryness. After purification by column chromatography, 40 g of colorless oil was obtained with a yield of 85%.
  • reaction solution was spin-dried, diluted with 500 mL of ethyl acetate, and washed twice with 500 mL of water.
  • the organic phase was dried over anhydrous sodium sulfate and then spin-dried. After purification by column chromatography, 25 g of colorless oil was obtained with a yield of 87%.
  • reaction solution was filtered through diatomaceous earth and spun to dryness. After purification by column chromatography, 2.8 g of colorless oil was obtained, with a yield of 83.6%.
  • reaction solution was filtered and spun to dryness. After purification by column chromatography, 2 g of colorless oil was obtained with a yield of 77%.
  • reaction solution was filtered through diatomaceous earth and spun to dryness. After purification by column chromatography, 2 g of colorless oil was obtained with a yield of 55%.
  • reaction solution was spin-dried, diluted with 100 mL of ethyl acetate, and washed twice with 100 mL of water.
  • the organic phase was dried over anhydrous sodium sulfate and then spin-dried. After purification by column chromatography, 750 mg of colorless oil was obtained, with a yield of 57.8%.
  • reaction solution was filtered and spun to dryness. After purification by column chromatography, 1 g of colorless oil was obtained with a yield of 76%.
  • reaction solution was filtered through diatomaceous earth and spun to dryness. After purification by column chromatography, 2 g of colorless oil was obtained.
  • reaction solution was filtered and spun to dryness. After purification by column chromatography, 2 g of colorless oil was obtained with a yield of 80%.
  • reaction solution was filtered through diatomaceous earth and spun to dryness. After purification by column chromatography, 2 g of colorless oil was obtained with a yield of 55%.
  • reaction solution was spin-dried, diluted with 100 ml of ethyl acetate, and washed twice with 100 ml of water.
  • the organic phase was dried over anhydrous sodium sulfate and then spun-dried. After purification by column chromatography, 750 mg of colorless oil was obtained, with a yield of 57.8%.
  • reaction solution was filtered and spun to dryness. After purification by column chromatography, 1 g of colorless oil was obtained with a yield of 76%.
  • reaction solution was filtered through diatomaceous earth and spun to dryness. After purification by column chromatography, 2 g of colorless oil was obtained.
  • reaction solution was filtered and spun to dryness. After purification by column chromatography, 2 g of colorless oil was obtained with a yield of 80%.
  • reaction solution was filtered through diatomaceous earth and spun to dryness. After purification by column chromatography, 2 g of colorless oil was obtained with a yield of 55%.
  • reaction solution was spin-dried, diluted with 100 ml of ethyl acetate, and washed twice with 100 ml of water.
  • the organic phase was dried over anhydrous sodium sulfate and then spun-dried. After purification by column chromatography, 750 mg of colorless oil was obtained, with a yield of 57.8%.
  • reaction solution was filtered and spun to dryness. After purification by column chromatography, 1 g of colorless oil was obtained with a yield of 76%.
  • reaction solution was filtered through diatomaceous earth and spun to dryness. After purification by column chromatography, 2 g of colorless oil was obtained.
  • reaction solution was filtered and spun to dryness. After purification by column chromatography, 2 g of colorless oil was obtained with a yield of 80%.
  • reaction solution was filtered through diatomaceous earth and spun to dryness. After purification by column chromatography, 2 g of colorless oil was obtained.
  • reaction solution was spin-dried, diluted with 100 mL of ethyl acetate, and washed twice with 100 ml of water.
  • the organic phase was dried over anhydrous sodium sulfate and then spin-dried. After purification by column chromatography, 750 mg of colorless oil was obtained, with a yield of 57.8%.
  • reaction solution was filtered and spun to dryness. After purification by column chromatography, 1 g of colorless oil was obtained with a yield of 76%.
  • reaction solution was filtered through diatomaceous earth and spun to dryness. After purification by column chromatography, 2 g of colorless oil was obtained.
  • reaction solution was filtered and spun to dryness. After purification by column chromatography, 2 g of colorless oil was obtained with a yield of 80%.
  • reaction solution was filtered through diatomaceous earth and spun to dryness. After purification by column chromatography, 2 g of colorless oil was obtained with a yield of 55%.
  • reaction solution was spin-dried, diluted with 100 mL of ethyl acetate, and washed twice with 100 mL of water.
  • the organic phase was dried over anhydrous sodium sulfate and then spin-dried. After purification by column chromatography, 750 mg of colorless oil was obtained, with a yield of 55%.
  • reaction solution was filtered and spun to dryness. After purification by column chromatography, 1 g of colorless oil was obtained with a yield of 73%.
  • reaction solution was purified by column chromatography to obtain 1.4 g of product.
  • reaction solution was filtered, spun to dryness, and purified by column chromatography to obtain 900 g of colorless oil.
  • reaction solution was filtered through diatomaceous earth and spun to dryness. After purification by column chromatography, 1 g of colorless oil was obtained with a yield of 25%.
  • reaction solution was spin-dried, diluted with 100 mL of ethyl acetate, and washed twice with 100 mL of water.
  • the organic phase was dried over anhydrous sodium sulfate and then spin-dried. After purification by column chromatography, 300 mg of colorless oil was obtained, with a yield of 32.8%.
  • reaction solution was filtered and spun to dryness. After purification by column chromatography, 160 mg of colorless oil was obtained, with a yield of 76%.
  • reaction solution was filtered through diatomaceous earth and spun to dryness. After purification by column chromatography, 2.8 g of colorless oil was obtained.
  • reaction solution was filtered and spun to dryness. After purification by column chromatography, 2 g of colorless oil was obtained with a yield of 80%.
  • reaction solution was filtered through diatomaceous earth and spun to dryness. After purification by column chromatography, 1.2 g of colorless oil was obtained, with a yield of 65%.
  • reaction solution was spin-dried, diluted with 100 mL of ethyl acetate, and washed twice with 100 mL of water.
  • the organic phase was dried over anhydrous sodium sulfate and spin-dried. After purification by column chromatography, 1 g of colorless oil was obtained with a yield of 87.8%.
  • reaction solution was filtered and spun to dryness. After purification by column chromatography, 1 g of colorless oil was obtained with a yield of 76%.
  • reaction solution was filtered through diatomaceous earth and spun to dryness. After purification by column chromatography, 8.6 g of colorless oil was obtained, with a yield of 69%.
  • reaction solution was spin-dried, diluted with 500 mL of ethyl acetate, and washed twice with 500 mL of water.
  • the organic phase was dried with anhydrous sodium sulfate and then spin-dried. After purification by column chromatography, 5.6 g of colorless oil was obtained. Yield 69%.
  • reaction solution was filtered, spun to dryness, and purified by column chromatography to obtain 1.55 g of colorless oil. Yield 64%.
  • reaction solution was filtered through diatomaceous earth and spun to dryness. After purification by column chromatography, 9 g of colorless oil was obtained with a yield of 70%.
  • reaction solution was filtered, spun to dryness, and purified by column chromatography to obtain 1 g of colorless oil. Yield 53%.
  • reaction solution was filtered through diatomaceous earth and spun to dryness. After purification by column chromatography, 2.4 g of colorless oil was obtained.
  • reaction solution was filtered and spun to dryness. After purification by column chromatography, 1.5 g of colorless oil was obtained. The yield is 71%.
  • reaction solution was filtered through diatomaceous earth and spun to dryness. After purification by column chromatography, 8.8 g of colorless oil was obtained, with a yield of 63%.
  • reaction solution was spin-dried, diluted with 500 mL of ethyl acetate, and washed twice with 500 mL of water.
  • the organic phase was dried over anhydrous sodium sulfate and then spin-dried. After purification by column chromatography, 5.8 g of colorless oil was obtained. The yield is 70%.
  • reaction solution was filtered, spun to dryness, and purified by column chromatography to obtain 1 g of colorless oil. Yield 57%.
  • reaction solution was filtered through diatomaceous earth and spun to dryness. After purification by column chromatography, 2.6 g of colorless oil was obtained.
  • reaction solution was filtered and spun to dryness. After purification by column chromatography, 1.2 g of colorless oil was obtained. Yield 57%.
  • reaction solution was filtered through diatomaceous earth and spun to dryness. After purification by column chromatography, 9.3 g of colorless oil was obtained, with a yield of 62%.
  • reaction solution was spin-dried, diluted with 500 mL of ethyl acetate, and washed twice with 500 mL of water.
  • the organic phase was dried over anhydrous sodium sulfate and then spin-dried. After purification by column chromatography, 5.4 g of colorless oil was obtained. Yield 60%.
  • reaction solution was filtered, spun to dryness, and purified by column chromatography to obtain 1 g of colorless oil. Yield 57%.
  • reaction solution was filtered through diatomaceous earth and spun to dryness. After purification by column chromatography, 2.8 g of colorless oil was obtained.
  • reaction solution was filtered and spun to dryness. After purification by column chromatography, 1.2 g of colorless oil was obtained. Yield 57%.
  • reaction solution was filtered through diatomaceous earth and spun to dryness. After purification by column chromatography, 9 g of colorless oil was obtained with a yield of 61%.
  • reaction solution was spin-dried, diluted with 500 mL of ethyl acetate, and washed twice with 500 mL of water.
  • the organic phase was dried with anhydrous sodium sulfate and then spin-dried. After purification by column chromatography, 5 g of colorless oil was obtained. Yield 57%.
  • reaction solution was filtered, spun to dryness, and purified by column chromatography to obtain 1.1 g of colorless oil. Yield 57%.
  • a series of ionizable lipid compounds distearoylphosphatidylcholine (DSPC, Nippon Seika Co., Ltd., product number: S01005), cholesterol (Nihon Seika Co., Ltd., product number: O01001) and dimyristoylglycerol-poly Ethylene glycol 2000 (DMG-PEG2000, Guobang Pharmaceutical, product number: O02005) was dissolved in ethanol solution, and then mixed according to a certain molar ratio to prepare an ethanol solution of mixed lipids (total lipid concentration is 12.5mM).
  • Firefly luciferase (Fluc) mRNA or SARS-CoV-2 spike protein (Spike) mRNA were dissolved in 50mM citric acid Dilute in salt buffer (pH 4.0) to obtain an mRNA solution.
  • Lipid nanoparticles were prepared by using a microfluidic device to mix the ethanol solution of lipid and the mRNA solution at a flow rate of 12 mL/min and a volume ratio of 1:3, with a nitrogen-phosphorus ratio of ionizable lipid to mRNA of 6:1.
  • SM102 ionizable lipid (Zhejiang Shenzhou Pharmaceutical Co., Ltd., batch number: W211-YB211202) was used as a positive control. Its structure is as follows:
  • the molar ratio of ionizable lipid/DSPC/cholesterol/DMG-PEG2000 is 50:10:38.5:1.5.
  • Malvern Zetasizer Ultra dynamic light scattering method
  • PDI polydispersity index
  • the encapsulation rate is used to indicate whether the LNP can effectively encapsulate mRNA.
  • the encapsulation rate is higher than 70%, which means that the LNP can effectively encapsulate mRNA.
  • Loading mRNA; single and bright bands in the agarose gel electrophoresis diagram indicate that the mRNA structure is complete. Among them, the closer the PDI is to 0, the better, and the closer the encapsulation rate is to 100%, the better.
  • each LNP prepared in Example 23 is between 60 and 150 nm, the PDI is less than 0.2, and the LNP pKa corresponding to each ionizable lipid is greater than 5.5.
  • the mRNA bands of each mRNA-LNP corresponding to SM102, LQ001, LQ007, LQ013, LQ016, LQ025, and LQ017 are clear and bright after electrophoresis, indicating that they can effectively encapsulate mRNA and maintain the integrity of the mRNA structure; the LNP corresponding to LQ109 There are no bright bands in the channels, indicating that the integrity of the mRNA in this group is poor.
  • LNP preparations encapsulated Fluc mRNA
  • lipid nanoparticles were prepared with a nitrogen-phosphorus ratio of ionizable lipid to mRNA of 6:1, and a molar ratio of ionizable lipid/DSPC/cholesterol/DMG-PEG2000 of 50:10:38.5:1.5, with four The total number of moles of the components is 100%, and the molar percentages of each group of preparations are consistent.
  • the particle size of each group of LNP preparations prepared from the above different ionizable lipids ranges from 60 to 250 nm; the PDI is less than 0.25, specifically between 0.038 and 0.234; the encapsulation rate is higher than 70%.
  • PBS is a negative control
  • the chemiluminescence intensity readings corresponding to this group can be regarded as background readings.
  • the Fluc-mRNA corresponding to LQ001, LQ007, LQ013, LQ016 and LQ025 Both LNPs can effectively deliver Fluc mRNA into cells and express it.
  • the Spike-mRNA LNP preparations corresponding to LQ001, LQ007, LQ013, and LQ025 can effectively deliver Spike mRNA into cells and express it.
  • the Fluc-mRNA-LNP prepared in Example 23 was injected into 6-8 week old female Balb/C mice (Vital Lever) through the tail vein at a dose of 5 ⁇ g/mouse, and at a specific time point after administration (6, 24, 48, 72 hours in this example) D-luciferin potassium salt was injected intraperitoneally, and then the radiation intensity of the protein expressed by the mRNA carried by the LNP in the mouse was detected by the IVIS Spectrum small animal live imager (corresponding to (bioluminescence expression intensity), and the total luminescence intensity of the mouse liver in vivo was calculated.
  • 10 minutes after intraperitoneal injection of luciferin 10 minutes after intraperitoneal injection of luciferin, luciferase activity was measured through bioluminescence imaging, and Living Image software (PerkinElmer) counted the total luminescence intensity in the liver area.
  • the Fluc-mRNA LNP preparations corresponding to LQ001, LQ007, LQ013, LQ016 and LQ025 all have strong expression in mice, and can still be observed to be 10 times higher than the background level (PBS group) until 72h. luminous intensity above.
  • the Spike-mRNA-LNP preparation prepared in Example 23 was injected intramuscularly into 6-8 week old female Balb/C mice (Vital Lever) at a dose of 2 ⁇ g/mouse, and repeated 21 days after the first injection. Inject the corresponding mRNA-LNP preparation. And at a specific time point after the second administration (the data in this example is one week after the second administration), the whole blood of the mice was collected. Serum was separated from whole blood by centrifugation at 2000 g for 10 min at 4°C. The serum was inactivated in a water bath at 56°C for 30 min and stored at -80°C for analysis.
  • SARS-CoV-2 (2019-nCoV)) Spike S1+S2ECD-His Recombinant Protein (Yiqiao Shenzhou, Cat. No.: 40589-V08B1) to coat the antigen
  • SARS-CoV-2 (2019- nCoV) Spike Neutralizing Antibody Mouse Mab (Yiqiao Shenzhou, Cat. No.: 40591-MM43) as control, 2% BSA blocked, Peroxidase AffiniPure Goat Anti-Mouse IgG (H+L) (Jackson ImmunoResearch, Cat.
  • the total antibody titer of Spike protein in the animals in the SM102, LQ001, LQ013, LQ025, LQ007, LQ010, and LQ016 groups increased significantly, while there was no increase in the LQ002, LQ014, and LQ109 groups.
  • Example 26 Referring to the mouse in vivo test method of Example 26, all the LNP preparations prepared in Example 24 were injected into 6 to 8-week-old female Balb/C mice through the tail vein at a dose of 5 ⁇ g/mouse, and after administration D-fluorescein potassium salt was injected intraperitoneally at specific time points (4, 24, and 48 hours in this example), and then the total luminescence intensity of the liver region was calculated using the same method as in Example 26. The test results are shown in Figure 6. The experimental results showed that 5 groups of LNP preparations (LQ031, LQ067, LQ073, LQ076, and LQ077) had strong expression in mice, indicating their better in vivo delivery capabilities.

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Abstract

Disclosed are a nitrogen-containing chain compound, a preparation method therefor, a composition containing said compound, and a use thereof. Provided is a nitrogen-containing chain compound as represented by formula (I) or a pharmaceutically acceptable salt thereof. The nitrogen-containing chain compound represented by formula (I) can be used for preparing a lipid carrier. The prepared lipid carrier can encapsulate a nucleic acid drug, and may be used for delivering a nucleic acid prophylactic agent and/or therapeutic agent to mammalian cells and organs and producing an effect.

Description

含氮链状化合物、其制备方法、包含其的组合物和应用Nitrogen-containing chain compounds, methods for their preparation, compositions containing the same and applications
本申请要求申请日为2022/8/12的中国专利申请202210969100X的优先权。本申请引用上述中国专利申请的全文。This application claims priority from Chinese patent application 202210969100X with a filing date of 2022/8/12. This application cites the full text of the above-mentioned Chinese patent application.
技术领域Technical field
本发明涉及含氮链状化合物、其制备方法、包含其的组合物和应用。The present invention relates to nitrogen-containing chain compounds, methods for their preparation, compositions containing them and their uses.
背景技术Background technique
核酸药物为现今基础和应用研究的一个重要方向。核酸药物可用于对病毒及细菌感染性疾病、肿瘤、代谢性疾病等的预防和/或治疗，其生产成本更低且周期更短，有利于快速开发个性化药物。然而，核酸为荷负电的大分子，难以透过细胞膜，同时核酸稳定性不佳，通过开发各种核酸包装和递送***可一定程度上克服核酸药物的不稳定性，提高其递送效率。Nucleic acid drugs are an important direction in current basic and applied research. Nucleic acid drugs can be used for the prevention and/or treatment of viral and bacterial infectious diseases, tumors, metabolic diseases, etc. Their production costs are lower and the cycle is shorter, which is conducive to the rapid development of personalized drugs. However, nucleic acids are negatively charged macromolecules that are difficult to penetrate through cell membranes. At the same time, nucleic acids have poor stability. By developing various nucleic acid packaging and delivery systems, the instability of nucleic acid drugs can be overcome to a certain extent and their delivery efficiency can be improved.
脂质纳米颗粒已被证明可用作递送生物活性物质(如小分子药物、蛋白质和核酸)进入细胞和/或细胞内区室的载体。通过设计及优化脂质纳米颗粒中的各组分的种类和用量，从而优化核酸药物递送***对于提高核酸药物预防及治疗的功效具重要意义，尤其是可用于递送RNA预防剂和/或治疗剂的脂质化合物以及相关的方法和组合物。Lipid nanoparticles have been shown to be useful as vehicles for delivering bioactive substances, such as small molecule drugs, proteins, and nucleic acids, into cells and/or intracellular compartments. Optimizing the nucleic acid drug delivery system by designing and optimizing the types and amounts of each component in lipid nanoparticles is of great significance to improve the efficacy of nucleic acid drug prevention and treatment, especially for the delivery of RNA preventive and/or therapeutic agents. Lipid compounds and related methods and compositions.
发明内容Contents of the invention
本发明旨在提供一种新的可用于递送核酸药物的可电离脂质化合物，增加可电离脂质化合物的种类及核酸预防剂和/或治疗剂递送载体的选择。为解决以上技术问题，本发明提供了一种含氮链状化合物、其制备方法、包含其的组合物和应用。本发明的组合物可用于高效递送核酸药物。The present invention aims to provide a new ionizable lipid compound that can be used to deliver nucleic acid drugs, and to increase the types of ionizable lipid compounds and the selection of nucleic acid preventive and/or therapeutic agent delivery carriers. In order to solve the above technical problems, the present invention provides a nitrogen-containing chain compound, its preparation method, compositions containing the same and applications. The composition of the present invention can be used to efficiently deliver nucleic acid drugs.
本发明的技术方案如下：The technical solution of the present invention is as follows:
本发明提供了一种如式I所示的含氮链状化合物或其药学上可接受的盐，
The present invention provides a nitrogen-containing chain compound shown in formula I or a pharmaceutically acceptable salt thereof,
X为C1-C6的亚烷基或C3-C6的亚环烷基；X is a C 1 -C 6 alkylene group or a C 3 -C 6 cycloalkylene group;
Y为未取代的或被1个、2个或3个Y1-1取代C1-C10的亚烷基；Y is an unsubstituted or C 1 -C 10 alkylene group substituted by 1, 2 or 3 Y 1-1 ;
各个Y1-1独立地为羟基、卤素或C1-C6的烷基；Each Y 1-1 is independently hydroxyl, halogen or C 1 -C 6 alkyl;
Z为未取代的或被1个、2个或3个Z1-1取代C1-C10的亚烷基；Z is an unsubstituted or C 1 -C 10 alkylene group substituted by 1, 2 or 3 Z 1-1 ;
各个Z1-1独立地为羟基、卤素或C1-C6的烷基； Each Z 1-1 is independently hydroxyl, halogen or C 1 -C 6 alkyl;
A1和A2独立地为 A 1 and A 2 are independently
R1为未取代的或被1个、2个或3个R1-1取代的C1-C20的烷基或未取代的或被1个、2个或3个R1-2取代的C2-C20的烯基；R 1 is a C 1 -C 20 alkyl group that is unsubstituted or substituted by 1, 2 or 3 R 1-1 or unsubstituted or substituted by 1, 2 or 3 R 1-2 C 2 -C 20 alkenyl;
各个R1-1独立地为羟基、C1-C10的烷基、未取代的或被1个、2个或3个R1-1-4取代的C3-C15的环烷基、 Each R 1-1 is independently a hydroxyl group, a C 1 -C 10 alkyl group, a C 3 -C 15 cycloalkyl group that is unsubstituted or substituted by 1, 2 or 3 R 1-1-4 ,
R1-1-1为C1-C6的烷基；R1-1-2为C1-C6的烷基；R1-1-3为C1-C6的烷基；各个R1-1-4独立地为C1-C6的烷基；R 1-1-1 is a C 1 -C 6 alkyl group; R 1-1-2 is a C 1 -C 6 alkyl group; R 1-1-3 is a C 1 -C 6 alkyl group; each R 1-1-4 is independently a C 1 -C 6 alkyl group;
各个R1-2独立地为羟基或C1-C10的烷基；Each R 1-2 is independently a hydroxyl group or a C 1 -C 10 alkyl group;
R2为未取代的或被1个、2个或3个R2-1取代的C1-C20的烷基或未取代的或被1个、2个或3个R2-2取代的C2-C20的烯基；R 2 is a C 1 -C 20 alkyl group that is unsubstituted or substituted by 1, 2 or 3 R 2-1 or unsubstituted or substituted by 1, 2 or 3 R 2-2 C 2 -C 20 alkenyl;
各个R2-1独立地为羟基、C1-C10的烷基、未取代的或被1个、2个或3个R2-1-4取代的C3-C15的环烷基、 Each R 2-1 is independently a hydroxyl group, a C 1 -C 10 alkyl group, a C 3 -C 15 cycloalkyl group that is unsubstituted or substituted by 1, 2 or 3 R 2-1-4 ,
R2-1-1为C1-C6的烷基；R2-1-2为C1-C6的烷基；R2-1-3为C1-C6的烷基；各个R2-1-4独立地为C1-C6的烷基；R 2-1-1 is a C 1 -C 6 alkyl group; R 2-1-2 is a C 1 -C 6 alkyl group; R 2-1-3 is a C 1 -C 6 alkyl group; each R 2-1-4 is independently a C 1 -C 6 alkyl group;
各个R2-2独立地为羟基或C1-C10的烷基；Each R 2-2 is independently a hydroxyl group or a C 1 -C 10 alkyl group;
当X为C1-C6的亚烷基时，R1为被1个、2个或3个羟基取代的C1-C20的烷基、未取代的或被1个、2个或3个R1-1-4取代的环戊烷、未取代的或被1个、2个或3个R1-1-4取代的环12烷、 When X is a C 1 -C 6 alkylene group, R 1 is a C 1 -C 20 alkyl group substituted by 1, 2 or 3 hydroxyl groups, unsubstituted, or Cyclopentane substituted with R 1-1-4 , cyclopentane unsubstituted or substituted by 1, 2 or 3 R 1-1-4 ,
某一优选方案中，如式I所示的含氮链状化合物或其药学上可接受的盐中，某些基团的定义可如下所述，其他基团的定义可如其他任一方案所述(以下简称“某一优选方案中”)：X中，所述C1-C6的亚烷基可为C1-C4的亚烷基，优选 例如 In a preferred embodiment, in the nitrogen-containing chain compound shown in Formula I or a pharmaceutically acceptable salt thereof, some groups can be defined as follows, and other groups can be defined as in any other embodiment. (hereinafter referred to as "in a preferred embodiment"): in X, the C 1 -C 6 alkylene group may be a C 1 -C 4 alkylene group, preferably For example
某一优选方案中，X中，所述C3-C6的亚环烷基可为 例如 In a certain preferred embodiment, in X, the C 3 -C 6 cycloalkylene group can be For example
某一优选方案中，Y中，所述C1-C10的亚烷基可为C4-C6的亚烷基，优选 例如 In a preferred embodiment, in Y, the C 1 -C 10 alkylene group may be a C 4 -C 6 alkylene group, preferably For example
某一优选方案中，各个Y1-1中，所述卤素可为氟、氯、溴或碘。In a certain preferred embodiment, in each Y 1-1 , the halogen can be fluorine, chlorine, bromine or iodine.
某一优选方案中，各个Y1-1中，所述C1-C6的亚烷基可为 In a certain preferred embodiment, in each Y 1-1 , the C 1 -C 6 alkylene group can be
某一优选方案中，Z中，所述C1-C10的亚烷基可为C5-C8的亚烷基，优选 例如 In a certain preferred embodiment, in Z, the C 1 -C 10 alkylene group may be a C 5 -C 8 alkylene group, preferably For example
某一优选方案中，各个Z1-1中，所述卤素可为氟、氯、溴或碘。In a certain preferred embodiment, in each Z 1-1 , the halogen may be fluorine, chlorine, bromine or iodine.
某一优选方案中，各个Z1-1中，所述C1-C6的亚烷基可为 In a certain preferred embodiment, in each Z 1-1 , the C 1 -C 6 alkylene group can be
某一优选方案中，R1中，所述C1-C20的烷基可为C1-C15的烷基，优选为直链烷基，例如乙基、丙基、己基、辛基、壬基或十二烷基。In a preferred embodiment, in R 1 , the C 1 -C 20 alkyl group can be a C 1 -C 15 alkyl group, preferably a straight chain alkyl group, such as ethyl, propyl, hexyl, octyl, Nonyl or dodecyl.
某一优选方案中，R1中，所述C2-C20的烯基可为C6-C18的烯基，优选为直链烯基；所述C2-C20的烯基可含有1到4个双键。In a preferred embodiment, in R 1 , the C 2 -C 20 alkenyl group may be a C 6 -C 18 alkenyl group, preferably a linear alkenyl group; the C 2 -C 20 alkenyl group may contain 1 to 4 double bonds.
某一优选方案中，各个R1-1中，所述C1-C10的烷基可为C1-C4的烷基，优选甲基、乙基、正丙基、异丙基、正丁基、叔丁基、异丁基或仲丁基，例如甲基、乙基、异丙基、叔丁基或正丁基。In a certain preferred embodiment, in each R 1-1 , the C 1 -C 10 alkyl group can be a C 1 -C 4 alkyl group, preferably methyl, ethyl, n-propyl, isopropyl, n-propyl, etc. Butyl, tert-butyl, isobutyl or sec-butyl, for example methyl, ethyl, isopropyl, tert-butyl or n-butyl.
某一优选方案中，各个R1-1中，所述C3-C15的环烷基可为C3-C12的环烷基，优选C4-C8的环烷基或环12烷基，例如环戊基、环己基或环12烷基。In a certain preferred embodiment, in each R 1-1 , the C 3 -C 15 cycloalkyl group can be a C 3 -C 12 cycloalkyl group, preferably a C 4 -C 8 cycloalkyl group or a cycloalkyl group. group, such as cyclopentyl, cyclohexyl or cycloalkyl.
某一优选方案中，R1-1-1中，所述C1-C6的烷基可为C1-C4的烷基，优选甲基、乙基、正丙基、异丙基、正丁基、叔丁基、异丁基或仲丁基。In a preferred embodiment, in R 1-1-1 , the C 1 -C 6 alkyl group can be a C 1 -C 4 alkyl group, preferably methyl, ethyl, n-propyl, isopropyl, n-butyl, tert-butyl, iso-butyl or sec-butyl.
某一优选方案中，R1-1-2中，所述C1-C6的烷基可为C1-C4的烷基，优选甲基、乙基、正丙基、异丙基、正丁基、叔丁基、异丁基或仲丁基。In a preferred embodiment, in R 1-1-2 , the C 1 -C 6 alkyl group can be a C 1 -C 4 alkyl group, preferably methyl, ethyl, n-propyl, isopropyl, n-butyl, tert-butyl, iso-butyl or sec-butyl.
某一优选方案中，R1-1-3中，所述C1-C6的烷基可为C1-C4的烷基，优选甲基、乙基、正丙基、异丙基、正丁基、叔丁基、异丁基或仲丁基。In a preferred embodiment, in R 1-1-3 , the C 1 -C 6 alkyl group can be a C 1 -C 4 alkyl group, preferably methyl, ethyl, n-propyl, isopropyl, n-butyl, tert-butyl, iso-butyl or sec-butyl.
某一优选方案中，各个R1-1-4中，所述C1-C6的烷基可为C1-C4的烷基，优选甲基、乙基、正丙基、异丙基、正丁基、叔丁基、异丁基或仲丁基。In a certain preferred embodiment, in each R 1-1-4 , the C 1 -C 6 alkyl group can be a C 1 -C 4 alkyl group, preferably methyl, ethyl, n-propyl, isopropyl , n-butyl, tert-butyl, isobutyl or sec-butyl.
某一优选方案中，各个R1-2中，所述C1-C10的烷基可为C1-C6的烷基，优选甲基、乙基、正丙基、 异丙基、正丁基、叔丁基、异丁基或仲丁基。In a certain preferred embodiment, in each R 1-2 , the C 1 -C 10 alkyl group can be a C 1 -C 6 alkyl group, preferably methyl, ethyl, n-propyl, Isopropyl, n-butyl, tert-butyl, isobutyl or sec-butyl.
某一优选方案中，R2中，所述C1-C20的烷基可为C1-C15的烷基，优选为直链烷基，例如乙基、丙基、己基、辛基、壬基或十二烷基。In a certain preferred embodiment, in R 2 , the C 1 -C 20 alkyl group can be a C 1 -C 15 alkyl group, preferably a straight chain alkyl group, such as ethyl, propyl, hexyl, octyl, Nonyl or dodecyl.
某一优选方案中，R2中，所述C2-C20的烯基可为C6-C18的烯基，优选为直链烯基；所述C2-C20的烯基可含有1到4个双键。In a preferred embodiment, in R 2 , the C 2 -C 20 alkenyl group may be a C 6 -C 18 alkenyl group, preferably a linear alkenyl group; the C 2 -C 20 alkenyl group may contain 1 to 4 double bonds.
某一优选方案中，各个R2-1中，所述C1-C10的烷基可为C1-C8的烷基，优选甲基、乙基、正丙基、异丙基、正丁基、叔丁基、异丁基、仲丁基、正戊基、异戊基、正己基、异己基、正庚基、异庚基、正辛基或异辛基，例如甲基、乙基、异丙基、叔丁基、正丁基或正辛基。In a preferred embodiment, in each R 2-1 , the C 1 -C 10 alkyl group can be a C 1 -C 8 alkyl group, preferably methyl, ethyl, n-propyl, isopropyl, n-propyl, etc. Butyl, tert-butyl, isobutyl, sec-butyl, n-pentyl, isopentyl, n-hexyl, isohexyl, n-heptyl, isoheptyl, n-octyl or isooctyl, such as methyl, ethyl base, isopropyl, tert-butyl, n-butyl or n-octyl.
某一优选方案中，各个R2-1中，所述C3-C15的环烷基可为C3-C12的环烷基，优选C4-C8的环烷基或环12烷基，例如环戊基、环己基或环12烷基。In a preferred embodiment, in each R 2-1 , the C 3 -C 15 cycloalkyl group can be a C 3 -C 12 cycloalkyl group, preferably a C 4 -C 8 cycloalkyl group or a cycloalkyl group. group, such as cyclopentyl, cyclohexyl or cycloalkyl.
某一优选方案中，R2-1-1中，所述C1-C6的烷基可为C1-C4的烷基，优选甲基、乙基、正丙基、异丙基、正丁基、叔丁基、异丁基或仲丁基。In a preferred embodiment, in R 2-1-1 , the C 1 -C 6 alkyl group can be a C 1 -C 4 alkyl group, preferably methyl, ethyl, n-propyl, isopropyl, n-butyl, tert-butyl, iso-butyl or sec-butyl.
某一优选方案中，R2-1-2中，所述C1-C6的烷基可为C1-C4的烷基，优选甲基、乙基、正丙基、异丙基、正丁基、叔丁基、异丁基或仲丁基。In a preferred embodiment, in R 2-1-2 , the C 1 -C 6 alkyl group can be a C 1 -C 4 alkyl group, preferably methyl, ethyl, n-propyl, isopropyl, n-butyl, tert-butyl, iso-butyl or sec-butyl.
某一优选方案中，R2-1-3中，所述C1-C6的烷基可为C1-C4的烷基，优选甲基、乙基、正丙基、异丙基、正丁基、叔丁基、异丁基或仲丁基。In a preferred embodiment, in R 2-1-3 , the C 1 -C 6 alkyl group can be a C 1 -C 4 alkyl group, preferably methyl, ethyl, n-propyl, isopropyl, n-butyl, tert-butyl, iso-butyl or sec-butyl.
某一优选方案中，各个R2-1-4中，所述C1-C6的烷基可为C1-C4的烷基，优选甲基、乙基、正丙基、异丙基、正丁基、叔丁基、异丁基或仲丁基。In a preferred embodiment, in each R 2-1-4 , the C 1 -C 6 alkyl group can be a C 1 -C 4 alkyl group, preferably methyl, ethyl, n-propyl, isopropyl , n-butyl, tert-butyl, isobutyl or sec-butyl.
某一优选方案中，各个R2-2中，所述C1-C10的烷基可为C1-C6的烷基，优选甲基、乙基、正丙基、异丙基、正丁基、叔丁基、异丁基或仲丁基。In a preferred embodiment, in each R 2-2 , the C 1 -C 10 alkyl group can be a C 1 -C 6 alkyl group, preferably methyl, ethyl, n-propyl, isopropyl, n-propyl, etc. Butyl, tert-butyl, isobutyl or sec-butyl.
某一优选方案中，所述如式I所示的含氮链状化合物为如式Ia、Ib、Ic、Id或Ie所示的化合物 In a certain preferred embodiment, the nitrogen-containing chain compound represented by formula I is a compound represented by formula Ia, Ib, Ic, Id or Ie.
某一优选方案中，A1为其中a与R1相连，b与Y相连，或a与Y相连，b与R1相连。In a certain preferred solution, A 1 is where a is connected to R 1 and b is connected to Y, or a is connected to Y and b is connected to R 1 .
某一优选方案中，A2为其中a与R2相连，b与Z相连，或a与Z相连，b与相连R2。In a certain preferred solution, A 2 is Where a is connected to R 2 and b is connected to Z, or a is connected to Z and b is connected to R 2 .
某一优选方案中，R1与R2相同。In a preferred embodiment, R 1 and R 2 are the same.
某一优选方案中，X为C1-C4的亚烷基或C3-C6的亚环烷基；In a preferred embodiment, X is a C 1 -C 4 alkylene group or a C 3 -C 6 cycloalkylene group;
Y为C1-C10的亚烷基；Y is a C 1 -C 10 alkylene group;
Z为C1-C10的亚烷基；Z is a C 1 -C 10 alkylene group;
A1和A2独立地为 A 1 and A 2 are independently
R1为未取代的或被1个、2个或3个R1-1取代的C1-C20的烷基；R 1 is a C 1 -C 20 alkyl group that is unsubstituted or substituted by 1, 2 or 3 R 1-1 ;
各个R1-1独立地为羟基、C1-C10的烷基或未取代的或被1个、2个或3个R1-1-4取代的C3-C15的环烷基；Each R 1-1 is independently a hydroxyl group, a C 1 -C 10 alkyl group or a C 3 -C 15 cycloalkyl group that is unsubstituted or substituted by 1, 2 or 3 R 1-1-4 ;
各个R1-1-4独立地为C1-C6的烷基；Each R 1-1-4 is independently a C 1 -C 6 alkyl group;
R2为未取代的或被1个、2个或3个R2-1取代的C1-C20的烷基；R 2 is a C 1 -C 20 alkyl group that is unsubstituted or substituted by 1, 2 or 3 R 2-1 ;
各个R2-1独立地为羟基、C1-C10的烷基或未取代的或被1个、2个或3个R2-1-4取代的C3-C15的环烷基；Each R 2-1 is independently a hydroxyl group, a C 1 -C 10 alkyl group or a C 3 -C 15 cycloalkyl group that is unsubstituted or substituted by 1, 2 or 3 R 2-1-4 ;
各个R2-1-4独立地为C1-C6的烷基。Each R 2-1-4 is independently a C 1 -C 6 alkyl group.
某一优选方案中，X为C1-C4的亚烷基或C3-C6的亚环烷基；In a preferred embodiment, X is a C 1 -C 4 alkylene group or a C 3 -C 6 cycloalkylene group;
Y为C4-C8的亚烷基；Y is a C 4 -C 8 alkylene group;
Z为C4-C8的亚烷基；Z is a C 4 -C 8 alkylene group;
A1为其中a与R1相连，b与Y相连；A 1 is Where a is connected to R 1 , b is connected to Y;
A2为其中a与R2相连，b与Z相连；A 2 is Among them, a is connected to R 2 , and b is connected to Z;
R1为未取代的或被1个、2个或3个R1-1取代的C1-C20的烷基；R 1 is a C 1 -C 20 alkyl group that is unsubstituted or substituted by 1, 2 or 3 R 1-1 ;
各个R1-1独立地为羟基、C1-C6的烷基或未取代的或被1个、2个或3个R1-1-4取代的C3-C6的环烷基； Each R 1-1 is independently a hydroxyl group, a C 1 -C 6 alkyl group or a C 3 -C 6 cycloalkyl group that is unsubstituted or substituted by 1, 2 or 3 R 1-1-4 ;
各个R1-1-4独立地为C1-C6的烷基；Each R 1-1-4 is independently a C 1 -C 6 alkyl group;
R2为未取代的或被1个、2个或3个R2-1取代的C1-C20的烷基；R 2 is a C 1 -C 20 alkyl group that is unsubstituted or substituted by 1, 2 or 3 R 2-1 ;
各个R2-1独立地为羟基、C1-C6的烷基或未取代的或被1个、2个或3个R2-1-4取代的C3-C6的环烷基；Each R 2-1 is independently a hydroxyl group, a C 1 -C 6 alkyl group or a C 3 -C 6 cycloalkyl group that is unsubstituted or substituted by 1, 2 or 3 R 2-1-4 ;
各个R2-1-4独立地为C1-C6的烷基；Each R 2-1-4 is independently a C 1 -C 6 alkyl group;
当X为C1-C4的亚烷基时，R1为被1个、2个或3个羟基取代的C1-C20的烷基、未取代的或被1个、2个或3个R1-1-4取代的环戊烷、 When X is a C 1 -C 4 alkylene group, R 1 is a C 1 -C 20 alkyl group substituted by 1, 2 or 3 hydroxyl groups, unsubstituted, or R 1-1-4 substituted cyclopentane,
某一优选方案中，X为C1-C4的亚烷基；In a certain preferred embodiment, X is a C 1 -C 4 alkylene group;
Y为C4-C8的亚烷基；Y is a C 4 -C 8 alkylene group;
Z为C4-C8的亚烷基；Z is a C 4 -C 8 alkylene group;
A1为其中a与R1相连，b与Y相连；A 1 is Where a is connected to R 1 , b is connected to Y;
A2为其中a与R2相连，b与Z相连；A 2 is Among them, a is connected to R 2 , and b is connected to Z;
R1为被1个、2个或3个羟基或环戊烷取代的C1-C15的烷基、 R 1 is a C 1 -C 15 alkyl group substituted by 1, 2 or 3 hydroxyl groups or cyclopentane,
R2为被1个、2个或3个R2-1取代的C1-C15的烷基；R 2 is a C 1 -C 15 alkyl group substituted by 1, 2 or 3 R 2-1 ;
各个R2-1独立地为C1-C10的烷基。Each R 2-1 is independently a C 1 -C 10 alkyl group.
某一优选方案中，X为C1-C4的亚烷基或C3-C6的亚环烷基；In a preferred embodiment, X is a C 1 -C 4 alkylene group or a C 3 -C 6 cycloalkylene group;
Y为C4-C8的亚烷基；Y is a C 4 -C 8 alkylene group;
Z为C4-C8的亚烷基；Z is a C 4 -C 8 alkylene group;
A1为其中a与R1相连，b与Y相连；A 1 is Where a is connected to R 1 , b is connected to Y;
A2为其中a与R2相连，b与Z相连； A 2 is Among them, a is connected to R 2 , and b is connected to Z;
R1为未取代的或被1个、2个或3个R1-1取代的C1-C20的烷基；R 1 is a C 1 -C 20 alkyl group that is unsubstituted or substituted by 1, 2 or 3 R 1-1 ;
各个R1-1独立地为羟基、C1-C6的烷基或未取代的或被1个、2个或3个R1-1-4取代的C3-C6的环烷基；Each R 1-1 is independently a hydroxyl group, a C 1 -C 6 alkyl group or a C 3 -C 6 cycloalkyl group that is unsubstituted or substituted by 1, 2 or 3 R 1-1-4 ;
各个R1-1-4独立地为C1-C6的烷基；Each R 1-1-4 is independently a C 1 -C 6 alkyl group;
R2为未取代的或被1个、2个或3个R2-1取代的C1-C20的烷基；R 2 is a C 1 -C 20 alkyl group that is unsubstituted or substituted by 1, 2 or 3 R 2-1 ;
各个R2-1独立地为羟基、C1-C8的烷基或未取代的或被1个、2个或3个R2-1-4取代的C3-C6的环烷基；Each R 2-1 is independently a hydroxyl group, a C 1 -C 8 alkyl group or a C 3 -C 6 cycloalkyl group that is unsubstituted or substituted by 1, 2 or 3 R 2-1-4 ;
各个R2-1-4独立地为C1-C6的烷基；Each R 2-1-4 is independently a C 1 -C 6 alkyl group;
当X为C1-C4的亚烷基时，R1为被1个、2个或3个羟基取代的C1-C20的烷基、未取代的或被1个、2个或3个R1-1-4取代的环戊烷、 When X is a C 1 -C 4 alkylene group, R 1 is a C 1 -C 20 alkyl group substituted by 1, 2 or 3 hydroxyl groups, unsubstituted, or R 1-1-4 substituted cyclopentane,
某一优选方案中，X为C1-C4的亚烷基；In a certain preferred embodiment, X is a C 1 -C 4 alkylene group;
Y为C4-C8的亚烷基；Y is a C 4 -C 8 alkylene group;
Z为C4-C8的亚烷基；Z is a C 4 -C 8 alkylene group;
A1为其中a与R1相连，b与Y相连；A 1 is Where a is connected to R 1 , b is connected to Y;
A2为其中a与R2相连，b与Z相连；A 2 is Among them, a is connected to R 2 , and b is connected to Z;
R1为被1个、2个或3个羟基或环戊烷取代的C1-C15的烷基、 R 1 is a C 1 -C 15 alkyl group substituted by 1, 2 or 3 hydroxyl groups or cyclopentane,
R2为被1个、2个或3个R2-1取代的C1-C15的烷基；R 2 is a C 1 -C 15 alkyl group substituted by 1, 2 or 3 R 2-1 ;
各个R2-1独立地为C1-C10的烷基或未取代的或被1个、2个或3个R1-1-4取代的C3-C6的环烷基； Each R 2-1 is independently a C 1 -C 10 alkyl group or a C 3 -C 6 cycloalkyl group that is unsubstituted or substituted by 1, 2 or 3 R 1-1-4 ;
各个R2-1-4独立地为C1-C6的烷基。Each R 2-1-4 is independently a C 1 -C 6 alkyl group.
某一优选方案中，X为C1-C4的亚烷基或C3-C6的亚环烷基。In a certain preferred embodiment, X is a C 1 -C 4 alkylene group or a C 3 -C 6 cycloalkylene group.
某一优选方案中，Y为C1-C10的亚烷基。In a certain preferred embodiment, Y is a C 1 -C 10 alkylene group.
某一优选方案中，Z为C1-C10的亚烷基。In a certain preferred embodiment, Z is a C 1 -C 10 alkylene group.
某一优选方案中，R1为未取代的或被1个、2个或3个R1-1取代的C1-C20的烷基。In a certain preferred embodiment, R 1 is a C 1 -C 20 alkyl group that is unsubstituted or substituted by 1, 2 or 3 R 1-1 .
某一优选方案中，各个R1-1独立地为羟基、C1-C10的烷基或未取代的或被1个、2个或3个R1-1- 4取代的C3-C15的环烷基。In a certain preferred embodiment, each R 1-1 is independently a hydroxyl group, a C 1 -C 10 alkyl group or an unsubstituted C 3 -C substituted by 1, 2 or 3 R 1-1-4 15 cycloalkyl.
某一优选方案中，R2为未取代的或被1个、2个或3个R2-1取代的C1-C20的烷基。In a certain preferred embodiment, R 2 is a C 1 -C 20 alkyl group that is unsubstituted or substituted by 1, 2 or 3 R 2-1 .
某一优选方案中，各个R2-1独立地为羟基、C1-C10的烷基或未取代的或被1个、2个或3个R2-1- 4取代的C3-C15的环烷基。In a certain preferred embodiment, each R 2-1 is independently a hydroxyl group, a C 1 -C 10 alkyl group or an unsubstituted C 3 -C substituted by 1, 2 or 3 R 2-1-4 15 cycloalkyl.
某一优选方案中，Y为C4-C8的亚烷基。In a certain preferred embodiment, Y is a C 4 -C 8 alkylene group.
某一优选方案中，Z为C4-C8的亚烷基。In a certain preferred embodiment, Z is a C 4 -C 8 alkylene group.
某一优选方案中，A1为其中a与R1相连，b与Y相连。In a certain preferred solution, A 1 is Where a is connected to R 1 and b is connected to Y.
某一优选方案中，A2为其中a与R2相连，b与Z相连。In a certain preferred solution, A 2 is Where a is connected to R 2 and b is connected to Z.
某一优选方案中，各个R1-1独立地为羟基、C1-C6的烷基或未取代的或被1个、2个或3个R1-1-4取代的C3-C6的环烷基。In a certain preferred embodiment, each R 1-1 is independently a hydroxyl group, a C 1 -C 6 alkyl group or an unsubstituted C 3 -C substituted by 1, 2 or 3 R 1-1-4 6 cycloalkyl.
某一优选方案中，各个R2-1独立地为羟基、C1-C6的烷基或未取代的或被1个、2个或3个R2-1-4取代的C3-C6的环烷基。In a certain preferred embodiment, each R 2-1 is independently a hydroxyl group, a C 1 -C 6 alkyl group or an unsubstituted C 3 -C substituted by 1, 2 or 3 R 2-1-4 6 cycloalkyl.
某一优选方案中，当X为C1-C4的亚烷基时，R1为被1个、2个或3个羟基取代的C1-C20的烷基、未取代的或被1个、2个或3个R1-1-4取代的环戊烷、 In a certain preferred embodiment, when X is a C 1 -C 4 alkylene group, R 1 is a C 1 -C 20 alkyl group substituted by 1, 2 or 3 hydroxyl groups, unsubstituted or One, two or three R 1-1-4 substituted cyclopentane,
某一优选方案中，X为 In a certain preferred solution, X is
某一优选方案中，Y为 In a certain preferred solution, Y is
某一优选方案中，Z为 In a certain preferred solution, Z is
某一优选方案中，R1为 In a certain preferred solution, R 1 is
某一优选方案中，R2为 In a certain preferred solution, R 2 is
某一优选方案中，所述如式I所示的含氮链状化合物为如下任一化合物：










In a certain preferred embodiment, the nitrogen-containing chain compound shown in Formula I is any of the following compounds:










本发明还提供一种如式I所示的含氮链状化合物的制备方法，其包括如下步骤：溶剂中，在碱和碘盐存在下，如式I-1所示的化合物与如式I-2所示的化合物进行如下式所示的偶联反应，即可；
The invention also provides a method for preparing a nitrogen-containing chain compound represented by formula I, which includes the following steps: in a solvent, in the presence of a base and an iodide salt, a compound represented by formula I-1 is mixed with a compound represented by formula I The compound represented by -2 can undergo a coupling reaction as shown in the following formula;
M为卤素，X、Y、Z、A1、A2、R1和R2如前所述。M is halogen, and X, Y, Z, A 1 , A 2 , R 1 and R 2 are as described above.
所述偶联反应中，所述卤素可为氟、氯、溴或碘，例如溴。In the coupling reaction, the halogen may be fluorine, chlorine, bromine or iodine, such as bromine.
所述偶联反应中，所述如式I-1所示的化合物与所述如式I-2所示的化合物的摩尔比可为1:(1-2)，例1:1.2。In the coupling reaction, the molar ratio of the compound represented by Formula I-1 to the compound represented by Formula I-2 may be 1:(1-2), Example 1:1.2.
所述偶联反应中，所述碱为本领域常规碱。所述碱可为碱式碳酸盐(盐中阳离子为碱金属离子，阴离子为碳酸根)，例如K2CO3。In the coupling reaction, the base is a conventional base in this field. The base may be a basic carbonate (the cation in the salt is an alkali metal ion and the anion is carbonate), such as K 2 CO 3 .
所述偶联反应中，所述如式I-1所示的化合物与所述碱的摩尔比可为1:(1-5)，例如1:3.5或1:2。In the coupling reaction, the molar ratio of the compound represented by Formula I-1 to the base may be 1:(1-5), such as 1:3.5 or 1:2.
所述偶联反应中，所述碘盐为本领域常规碘盐。所述碘盐可为碱式碘盐，例如KI。In the coupling reaction, the iodized salt is a conventional iodized salt in this field. The iodized salt may be a basic iodized salt, such as KI.
所述偶联反应中，所述如式I-1所示的化合物与所述碘盐的摩尔比可为1:(1-2)，例如1:1.2或1:1。In the coupling reaction, the molar ratio of the compound represented by Formula I-1 to the iodide salt may be 1:(1-2), such as 1:1.2 or 1:1.
所述偶联反应中，所述溶剂为本领域常规溶剂，所述溶剂可为醚类溶剂或/和腈类溶剂。所述醚类溶剂可为甲基叔丁基醚。所述腈类溶剂可为乙腈。所述腈类溶剂与醚类溶剂的体积比可为1:1。In the coupling reaction, the solvent is a conventional solvent in this field, and the solvent can be an ether solvent or/and a nitrile solvent. The ether solvent may be methyl tert-butyl ether. The nitrile solvent may be acetonitrile. The volume ratio of the nitrile solvent to the ether solvent can be 1:1.
所述偶联反应中，所述如式I-1所示的化合物与所述溶剂的质量体积比可为10mg/mL-65mg/mL，例如15mg/mL、25mg/mL、40mg/mL、58.5mg/mL、20mg/L、22mg/L或34mg/L。In the coupling reaction, the mass-to-volume ratio of the compound represented by Formula I-1 to the solvent may be 10 mg/mL-65 mg/mL, such as 15 mg/mL, 25 mg/mL, 40 mg/mL, 58.5 mg/mL, 20mg/L, 22mg/L or 34mg/L.
所述偶联反应中，所述偶联反应的反应温度为本领域常规反应温度，优选为70℃-90℃，例如80℃。In the coupling reaction, the reaction temperature of the coupling reaction is a conventional reaction temperature in this field, preferably 70°C to 90°C, such as 80°C.
本发明还提供一种脂质载体，其包括物质Z，所述物质Z为如前所述的如式I所示化合物或其药学可接受的盐。The present invention also provides a lipid carrier, which includes substance Z, which is a compound represented by Formula I as described above or a pharmaceutically acceptable salt thereof.
某一优选方案中，所述脂质载体还包括稀释剂。In a preferred embodiment, the lipid carrier further includes a diluent.
某一优选方案中，所述稀释剂可为磷酸盐缓冲液或Tris缓冲液等。In a preferred embodiment, the diluent may be phosphate buffer or Tris buffer, etc.
某一优选方案中，所述脂质载体还包括磷脂。In a preferred embodiment, the lipid carrier further includes phospholipids.
某一优选方案中，所述磷脂可为本领域常规磷脂，其为两性辅助性分子，有助于脂质颗粒和细胞膜的融合。所述磷脂可为具有带电极性端和脂肪链非极性端的磷脂类分子，例如二硬脂酰基磷脂酰胆碱(DSPC)、二肉豆蔻酰磷酸胆碱(DMPC)、二油酰磷酸胆碱(DOPC)、棕榈酰磷酸胆碱(DPPC)、1,2-二硬脂酰磷酸胆碱(DSPC)、二十一烷酰磷酸胆碱(DUPC)或棕榈酰磷酸胆碱(POPC)等。In a preferred embodiment, the phospholipid can be a conventional phospholipid in the art, which is an amphoteric auxiliary molecule that contributes to the fusion of lipid particles and cell membranes. The phospholipid may be a phospholipid molecule with a charged polar end and a non-polar end of the fatty chain, such as distearoyl phosphatidylcholine (DSPC), dimyristoyl phosphocholine (DMPC), dioleoyl phosphocholine alkali (DOPC), palmitoyl phosphocholine (DPPC), 1,2-distearoyl phosphocholine (DSPC), undecanoyl phosphocholine (DUPC) or palmitoyl phosphocholine (POPC), etc. .
某一优选方案中，所述脂质载体还包括PEG脂质(聚乙二醇修饰的脂质)。In a certain preferred embodiment, the lipid carrier also includes PEG lipid (polyethylene glycol modified lipid).
某一优选方案中，所述PEG脂质可为具有聚乙二醇亲水端修饰的脂质分子。所述PEG脂质优选选自PEG修饰的磷脂酰乙醇胺、PEG修饰的磷脂酸、PEG修饰的神经酰胺、PEG修饰的二烷基胺、PEG修饰的二酰基甘油和PEG修饰的二烷基甘油中的一种或多种，例如PEG修饰的二肉豆蔻酰甘油(DMG-PEG2000)等。In a preferred embodiment, the PEG lipid may be a lipid molecule modified with a polyethylene glycol hydrophilic end. The PEG lipid is preferably selected from the group consisting of PEG-modified phosphatidylethanolamine, PEG-modified phosphatidic acid, PEG-modified ceramide, PEG-modified dialkylamine, PEG-modified diacylglycerol and PEG-modified dialkylglycerol. One or more, such as PEG-modified dimyristoylglycerol (DMG-PEG2000), etc.
某一优选方案中，所述脂质载体还包括甾醇。 In a preferred embodiment, the lipid carrier further includes sterols.
某一优选方案中，所述甾醇可为本领域常规甾醇，所述甾醇包括动物性、植物性或菌类甾醇。所述甾醇选自胆固醇、谷甾醇、麦角甾醇、菜油甾醇、豆甾醇、芸苔甾醇、番茄碱、熊果酸和α-生育酚中的一种或多种，例如胆固醇等。In a certain preferred embodiment, the sterols can be conventional sterols in this field, and the sterols include animal, plant or fungal sterols. The sterol is selected from one or more of cholesterol, sitosterol, ergosterol, campesterol, stigmasterol, brassinosterol, tomatine, ursolic acid and α-tocopherol, such as cholesterol and the like.
某一优选方案中，所述脂质载体中，所述物质Z与甾醇的摩尔比为0.5～5:1，优选为0.5～3:1，例如0.6～2:1。In a certain preferred embodiment, in the lipid carrier, the molar ratio of the substance Z to the sterol is 0.5-5:1, preferably 0.5-3:1, for example, 0.6-2:1.
某一优选方案中，所述脂质载体中，所述物质Z与磷脂的摩尔比为1～15:1，优选为2～8:1，例如3～6:1。In a certain preferred embodiment, in the lipid carrier, the molar ratio of the substance Z to the phospholipid is 1 to 15:1, preferably 2 to 8:1, such as 3 to 6:1.
某一优选方案中，所述脂质载体中，所述物质Z与PEG脂质的摩尔比为20～130:1，优选为20～80:1，例如20～40:1。In a certain preferred embodiment, in the lipid carrier, the molar ratio of the substance Z to the PEG lipid is 20 to 130:1, preferably 20 to 80:1, such as 20 to 40:1.
某一优选方案中，所述物质Z的摩尔含量约为30mol％至60mol％。In a certain preferred embodiment, the molar content of substance Z is about 30 mol% to 60 mol%.
本发明中，摩尔含量的含义为某物质占脂质载体的总物质量的百分比，脂质载体中各组分的摩尔含量之和不超过100mol％。某一优选方案中，所述磷脂的摩尔含量约为0mol％至30mol％。In the present invention, the molar content means the percentage of a certain substance in the total mass of the lipid carrier, and the sum of the molar contents of each component in the lipid carrier does not exceed 100 mol%. In a certain preferred embodiment, the molar content of the phospholipid is about 0 mol% to 30 mol%.
某一优选方案中，所述甾醇的摩尔含量约为15mol％至55mol％。In a certain preferred embodiment, the molar content of the sterol is about 15 mol% to 55 mol%.
某一优选方案中，所述PEG脂质的摩尔含量约为0mol％至10mol％。In a certain preferred embodiment, the molar content of the PEG lipid is about 0 mol% to 10 mol%.
某一优选方案中，所述脂质载体由所述物质Z、所述稀释剂、所述磷脂、所述PEG脂质和所述甾醇组成。In a certain preferred embodiment, the lipid carrier consists of the substance Z, the diluent, the phospholipid, the PEG lipid and the sterol.
本发明还提供一种脂质纳米颗粒，其包括治疗剂和/或预防剂以及前述脂质载体。The present invention also provides a lipid nanoparticle, which includes a therapeutic agent and/or a preventive agent and the aforementioned lipid carrier.
某一优选方案中，所述治疗剂和/或预防剂可为一种或两种及以上核酸。所述核酸可为本领域常规核酸。所述治疗剂和/或预防剂可为单链脱氧核糖核酸(DNA)、双链DNA、小干扰RNA(siRNA)、不对称双链小干扰RNA(aiRNA)、微小RNA(miRNA)、小发夹RNA(shRNA)、环状RNA(circRNA)、转运RNA(tRNA)、信使RNA(mRNA)和本领域已知的其他形式的核酸分子，优选为mRNA，例如萤火虫荧光素酶(Fluc)mRNA或SARS-CoV-2刺突蛋白(Spike)mRNA。In a certain preferred embodiment, the therapeutic agent and/or preventive agent may be one or two or more nucleic acids. The nucleic acid may be a conventional nucleic acid in the art. The therapeutic and/or preventive agent may be single-stranded deoxyribonucleic acid (DNA), double-stranded DNA, small interfering RNA (siRNA), asymmetric double-stranded small interfering RNA (aiRNA), microRNA (miRNA), small RNA Clamp RNA (shRNA), circular RNA (circRNA), transfer RNA (tRNA), messenger RNA (mRNA) and other forms of nucleic acid molecules known in the art, preferably mRNA, such as firefly luciferase (Fluc) mRNA or SARS-CoV-2 spike protein (Spike) mRNA.
某一优选方案中，所述脂质纳米颗粒中氮磷比可为2:1～30:1，所述组合物的氮磷比是指一种或多种可电离脂质化合物中可电离氮原子的摩尔数与RNA中磷酸酯基的摩尔数的比率。优选为2:1～20:1，例如3:1～20:1，还例如3:1～16:1。In a certain preferred embodiment, the nitrogen to phosphorus ratio in the lipid nanoparticles can be 2:1 to 30:1, and the nitrogen to phosphorus ratio of the composition refers to the ionizable nitrogen in one or more ionizable lipid compounds. The ratio of the number of moles of atoms to the number of moles of phosphate groups in RNA. Preferably, it is 2:1 to 20:1, such as 3:1 to 20:1, and another example is 3:1 to 16:1.
某一优选方案中，所述脂质纳米颗粒中，所述脂质载体与治疗剂和/或预防剂的质量比可为3～80:1，优选为6～60:1。In a certain preferred embodiment, in the lipid nanoparticles, the mass ratio of the lipid carrier to the therapeutic agent and/or preventive agent can be 3 to 80:1, preferably 6 to 60:1.
某一方案中，所述脂质纳米颗粒的粒径(平均粒径)可为10～250nm，例如为40～250nm，进一步为50～250nm，或40～150nm，或60～150nm。In a certain solution, the particle size (average particle size) of the lipid nanoparticles can be 10 to 250 nm, for example, 40 to 250 nm, further 50 to 250 nm, or 40 to 150 nm, or 60 to 150 nm.
某一优选方案中，所述脂质纳米颗粒中，所述脂质载体包裹所述治疗剂和/或预防剂。In a preferred embodiment, in the lipid nanoparticles, the lipid carrier encapsulates the therapeutic agent and/or preventive agent.
本发明还提供一种组合物，其包括物质Z，所述物质Z为如前所述的如式I所示化合物或其药学可接受的盐。The present invention also provides a composition, which includes substance Z, which is a compound represented by Formula I as described above or a pharmaceutically acceptable salt thereof.
某一优选方案中，所述组合物还包括稀释剂、磷脂、PEG脂质、甾醇和治疗剂和/或预防剂中的一种或多种。 In a certain preferred embodiment, the composition further includes one or more of diluent, phospholipid, PEG lipid, sterol and therapeutic and/or preventive agent.
某一优选方案中，所述组合物中，所述稀释剂、磷脂、PEG脂质、甾醇和治疗剂和/或预防剂如前所述。In a certain preferred embodiment, in the composition, the diluent, phospholipid, PEG lipid, sterol and therapeutic agent and/or preventive agent are as described above.
某一优选方案中，所述组合物中，所述物质Z与所述稀释剂、磷脂、PEG脂质和甾醇中的一种或多种形成如前所述脂质载体。In a certain preferred embodiment, in the composition, the substance Z and one or more of the diluent, phospholipid, PEG lipid and sterol form a lipid carrier as described above.
某一优选方案中，所述组合物中，所述脂质载体与所述治疗剂和/或预防剂形成如前所述的脂质纳米颗粒。某一优选方案中，所述组合物中，所述治疗剂和/或预防剂的包封率为至少50％，优选为至少为70％。In a certain preferred embodiment, in the composition, the lipid carrier and the therapeutic agent and/or preventive agent form lipid nanoparticles as described above. In a certain preferred embodiment, the encapsulation rate of the therapeutic agent and/or preventive agent in the composition is at least 50%, preferably at least 70%.
某一优选方案中，所述组合物中，所述组合物的多分散指数不高于0.5，例如不高于0.3。In a certain preferred embodiment, in the composition, the polydispersity index of the composition is not higher than 0.5, for example, not higher than 0.3.
如无特别说明，本发明所用术语具有如下含义：Unless otherwise specified, the terms used in this invention have the following meanings:
术语“卤素”是指氟、氯、溴或碘。The term "halogen" refers to fluorine, chlorine, bromine or iodine.
术语“药学上可接受”是指相对无毒、安全、适合于患者使用。The term "pharmaceutically acceptable" means relatively nontoxic, safe, and suitable for use by patients.
术语“药学上可接受的盐”是指化合物与药学上可接受的酸或碱反应得到的盐。当化合物中含有相对酸性的官能团时，可以通过在合适的惰性溶剂中用足量的药学上可接受的碱与化合物接触的方式获得碱加成盐。药学上可接受的碱加成盐包括但不限于：钠盐、钾盐、钙盐、铝盐、镁盐、铋盐、铵盐等。当化合物中含有相对碱性的官能团时，可以通过在合适的惰性溶剂中用足量的药学上可接受的酸与化合物接触的方式获得酸加成盐。药学上可接受的酸加成盐包括但不限于：盐酸盐、硫酸盐、甲磺酸盐等。具体可参见Handbook of Pharmaceutical Salts:Properties,Selection,and Use(P.Heinrich Stahl,Camille G.Wermuth,2011,2nd Revised Edition)。The term "pharmaceutically acceptable salt" refers to a salt obtained by reacting a compound with a pharmaceutically acceptable acid or base. When the compound contains relatively acidic functional groups, base addition salts can be obtained by contacting the compound with a sufficient amount of a pharmaceutically acceptable base in a suitable inert solvent. Pharmaceutically acceptable base addition salts include, but are not limited to, sodium salts, potassium salts, calcium salts, aluminum salts, magnesium salts, bismuth salts, ammonium salts, etc. When the compound contains relatively basic functional groups, acid addition salts can be obtained by contacting the compound with a sufficient amount of a pharmaceutically acceptable acid in a suitable inert solvent. Pharmaceutically acceptable acid addition salts include, but are not limited to: hydrochloride, sulfate, methanesulfonate, etc. For details, see Handbook of Pharmaceutical Salts: Properties, Selection, and Use (P.Heinrich Stahl, Camille G.Wermuth, 2011, 2nd Revised Edition).
结构片段中的是指该结构片段通过该位点与分子其余部分相连。例如，是指环己基。in structure fragment It means that the structural fragment is connected to the rest of the molecule through this site. For example, It refers to cyclohexyl.
基团末端的“-”是指该基团通过该位点与分子其余部分相连。例如，CH3-C(＝O)-是指乙酰基。The "-" at the end of a group means that the group is connected to the rest of the molecule through that site. For example, CH3 -C(=O)- refers to acetyl.
术语“烷基”是指具有指定碳原子数(例如，C1～C6)的、直链或支链的、饱和的一价烃基。烷基包括但不限于：甲基、乙基、正丙基、异丙基、正丁基、异丁基、仲丁基、叔丁基、正戊基、正己基等。The term "alkyl" refers to a linear or branched, saturated monovalent hydrocarbon group having a specified number of carbon atoms (eg, C 1 to C 6 ). Alkyl groups include, but are not limited to: methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, sec-butyl, tert-butyl, n-pentyl, n-hexyl, etc.
术语“亚烷基”为二价基团，其通过两个单键与分子其余部分相连，其余定义同术语“烷基”。The term "alkylene" is a divalent group connected to the rest of the molecule by two single bonds and is otherwise defined in the same manner as the term "alkyl".
术语“环烷基”是指具有指定的碳原子数(例如C3～C6)的、仅由碳原子组成的、饱和的单环环状基团。环烷基包括但不限于环丙基、环丁基、环戊基、环己基等。The term "cycloalkyl" refers to a saturated monocyclic cyclic group having a specified number of carbon atoms (eg, C 3 to C 6 ) and consisting only of carbon atoms. Cycloalkyl groups include, but are not limited to, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, and the like.
术语“烯基”是指包含一个或多个碳-碳双键(例如，1、2或3碳-碳双键)的直链或支链烃基团。The term "alkenyl" refers to a straight or branched chain hydrocarbon group containing one or more carbon-carbon double bonds (eg, 1, 2, or 3 carbon-carbon double bonds).
当任意变量(例如基团R1-1)在化合物的定义中多次出现时，它们的定义互相独立、互不影响。例如，被3个R1-1取代的C6～C10芳基是指C6～C10芳基会被3个R1-1取代，3个R1-1的定义互相独立的、互不影响。When any variable (such as the group R 1-1 ) appears multiple times in the definition of a compound, their definitions are independent of each other and do not affect each other. For example, a C 6 to C 10 aryl group substituted by 3 R 1-1 means that the C 6 to C 10 aryl group will be substituted by 3 R 1-1 . The definitions of the three R 1-1 are independent of each other. No impact.
在不违背本领域常识的基础上，上述各优选条件，可任意组合，即得本发明各较佳实例 On the basis of not violating common sense in the field, the above preferred conditions can be combined arbitrarily to obtain preferred examples of the present invention.
本发明所用试剂和原料均市售可得。The reagents and raw materials used in the present invention are all commercially available.
本发明的积极进步效果在于：本发明提供了一种如式I所示的含氮链状化合物，其结构新颖，其可用做制备脂质纳米颗粒。包含如式I所示的含氮链状化合物的脂质纳米颗粒具有较低的多分散指数，可以高效的转运mRNA。The positive and progressive effect of the present invention is that the present invention provides a nitrogen-containing chain compound shown in Formula I, which has a novel structure and can be used to prepare lipid nanoparticles. Lipid nanoparticles containing nitrogen-containing chain compounds as shown in Formula I have a low polydispersity index and can transport mRNA efficiently.
附图说明Description of drawings
图1为实施23中制备的各Fluc-mRNA LNP凝胶电泳图。Figure 1 shows the gel electrophoresis pattern of each Fluc-mRNA LNP prepared in Implementation 23.
图2为实施例25中293FT细胞与各Fluc-mRNA LNP共培养18-24h后的化学发光强度。Figure 2 shows the chemiluminescence intensity of 293FT cells in Example 25 after co-culture with each Fluc-mRNA LNP for 18-24 hours.
图3为实施例25中293FT细胞与各Spike-mRNA LNP共培养24h后的Spike蛋白表达情况，PBS组作为阴性对照。Figure 3 shows the Spike protein expression of 293FT cells and each Spike-mRNA LNP after co-culture for 24 hours in Example 25, and the PBS group was used as a negative control.
图4为实施例26中小鼠静脉注射给药各Fluc-mRNA LNP不同时间后的体内总生物发光量(n＝3)，PBS组做阴性对照(n＝2)。Figure 4 shows the total bioluminescence amount in mice after intravenous injection of each Fluc-mRNA LNP for different times in Example 26 (n=3), and the PBS group serves as a negative control (n=2).
图5为实施例26中小鼠肌内注射给药各Spike-mRNA LNP后的体内总抗体滴度(末次给药后1周，n＝8)。Figure 5 shows the total antibody titer in mice after intramuscular injection of each Spike-mRNA LNP in Example 26 (1 week after the last administration, n=8).
图6为实施例27中小鼠静脉注射给药各Fluc mRNA LNP不同时间后的体内总生物发光量(n＝3)。Figure 6 shows the total bioluminescence amount in mice after intravenous injection of each Fluc mRNA LNP for different times in Example 27 (n=3).
具体实施方式Detailed ways
下面通过实施例的方式进一步说明本发明，但并不因此将本发明限制在所述的实施例范围之中。下列实施例中未注明具体条件的实验方法，按照常规方法和条件，或按照商品说明书选择。The present invention is further described below by means of examples, but the present invention is not limited to the scope of the described examples. Experimental methods that do not indicate specific conditions in the following examples should be selected according to conventional methods and conditions, or according to product specifications.
本申请实施例的试剂来源如下：The sources of reagents in the examples of this application are as follows:
8-溴辛酸：购自江苏艾康，纯度：98％；8-Bromooctanoic acid: purchased from Jiangsu Aikang, purity: 98%;
十七烷-9-醇：购自北京福源坊科技有限公司，纯度：98％；Heptadecan-9-ol: purchased from Beijing Fuyuanfang Technology Co., Ltd., purity: 98%;
DCC：二环己基碳二亚胺，购自阿达玛斯试剂有限公司，货号012041444，纯度：RG，99％；DCC: dicyclohexylcarbodiimide, purchased from Adamas Reagent Co., Ltd., product number 012041444, purity: RG, 99%;
DMAP：4-二甲氨基吡啶，购自阿达玛斯试剂有限公司，货号：01271081，纯度：RG，99％；DMAP: 4-dimethylaminopyridine, purchased from Adamas Reagent Co., Ltd., product number: 01271081, purity: RG, 99%;
DCM：二氯甲烷，购自上海泰坦科技股份有限公司，货号：01111853，纯度：AR，≥99.5％；DCM: dichloromethane, purchased from Shanghai Titan Technology Co., Ltd., item number: 01111853, purity: AR, ≥99.5%;
乙醇胺：购自上海易恩化学技术有限公司，货号：R016710，纯度：AR，99％；Ethanolamine: purchased from Shanghai Yien Chemical Technology Co., Ltd., product number: R016710, purity: AR, 99%;
乙腈：购自上海泰坦科技股份有限公司，货号：01111797，纯度：AR，≥99.0％；Acetonitrile: purchased from Shanghai Titan Technology Co., Ltd., product number: 01111797, purity: AR, ≥99.0%;
6-溴己酸：购自阿达玛斯试剂有限公司，货号：01073739，纯度：RG，98％+；6-Bromohexanoic acid: purchased from Adamas Reagent Co., Ltd., item number: 01073739, purity: RG, 98%+;
3,7-二甲基-1-辛醇：购自安耐吉化学，货号：A040472，纯度：98％；3,7-Dimethyl-1-octanol: purchased from Anaiji Chemical, product number: A040472, purity: 98%;
K2CO3：购自上海易恩化学技术有限公司，货号：RH425011，纯度：AR，99％；K 2 CO 3 : purchased from Shanghai Yien Chemical Technology Co., Ltd., item number: RH425011, purity: AR, 99%;
KI：购自上海易恩化学技术有限公司，货号：RH432132，纯度：AR，99％；KI: purchased from Shanghai Yien Chemical Technology Co., Ltd., item number: RH432132, purity: AR, 99%;
甲基叔丁基醚：购自上海泰坦科技股份有限公司，货号：01030342，纯度：AR，≥99.0％；Methyl tert-butyl ether: purchased from Shanghai Titan Technology Co., Ltd., product number: 01030342, purity: AR, ≥99.0%;
2-环己基乙醇：购自上海阿拉丁生化科技股份有限公司，货号：C153306，纯度：GC，＞98.0％； 2-Cyclohexylethanol: purchased from Shanghai Aladdin Biochemical Technology Co., Ltd., product number: C153306, purity: GC, >98.0%;
异癸醇：购自上海阿拉丁生化科技股份有限公司，货号：I298943，纯度：98％；Isodecanol: purchased from Shanghai Aladdin Biochemical Technology Co., Ltd., product number: I298943, purity: 98%;
硅藻土：购自上海泰坦科技股份有限公司，货号：01589000，纯度：特优级，≥89.0％，200目；Diatomite: purchased from Shanghai Titan Technology Co., Ltd., item number: 01589000, purity: premium grade, ≥89.0%, 200 mesh;
乙酸乙酯：购自上海泰坦科技股份有限公司，货号：01153552，纯度：AR，≥99.5％；Ethyl acetate: purchased from Shanghai Titan Technology Co., Ltd., product number: 01153552, purity: AR, ≥99.5%;
3,5,5-三甲基-1-己醇：购自上海阿拉丁生化科技股份有限公司，货号：T162750，纯度：GC，＞85.0％；3,5,5-Trimethyl-1-hexanol: purchased from Shanghai Aladdin Biochemical Technology Co., Ltd., product number: T162750, purity: GC, >85.0%;
2-乙基己醇：购自上海易恩化学技术有限公司，货号：R016621，纯度：AR，99％；2-Ethylhexanol: purchased from Shanghai Yien Chemical Technology Co., Ltd., product number: R016621, purity: AR, 99%;
无水硫酸钠：购自上海泰坦科技股份有限公司，货号：01224581，纯度：AR，≥99.0％；Anhydrous sodium sulfate: purchased from Shanghai Titan Technology Co., Ltd., product number: 01224581, purity: AR, ≥99.0%;
三氯化铁：购自上海麦克林生化科技股份有限公司，货号：I811935，纯度：AR，99％；Ferric chloride: purchased from Shanghai McLean Biochemical Technology Co., Ltd., item number: I811935, purity: AR, 99%;
吡啶：购自上海麦克林生化科技股份有限公司，货号：P816288，纯度：AR，99％；Pyridine: purchased from Shanghai McLean Biochemical Technology Co., Ltd., product number: P816288, purity: AR, 99%;
1,2-环氧十二烷：购自上海麦克林生化科技股份有限公司，货号：E808874，纯度：95％；1,2-Epoxydodecane: purchased from Shanghai McLean Biochemical Technology Co., Ltd., product number: E808874, purity: 95%;
3-环戊基-1-丙醇：购自上海毕得医药科技股份有限公司，货号：BD86403，纯度：98％；3-Cyclopentyl-1-propanol: purchased from Shanghai Bid Pharmaceutical Technology Co., Ltd., product number: BD86403, purity: 98%;
环己基甲醇：购自上海易恩化学技术有限公司，货号：R014045，纯度：99％；Cyclohexylmethanol: purchased from Shanghai Yien Chemical Technology Co., Ltd., product number: R014045, purity: 99%;
异壬醇：购自广东翁江化学试剂有限公司；Isononyl alcohol: purchased from Guangdong Wengjiang Chemical Reagent Co., Ltd.;
8-甲基-1-癸醇：购自南京一览生物科技有限公司，纯度：97％；8-Methyl-1-decanol: purchased from Nanjing Yilan Biotechnology Co., Ltd., purity: 97%;
3-丁基-1-庚醇：购自苏州美诺医药科技有限公司，纯度：95％。3-Butyl-1-heptanol: purchased from Suzhou Meinuo Pharmaceutical Technology Co., Ltd., purity: 95%.
实施例1制备化合物LQ001Example 1 Preparation of Compound LQ001
步骤1：LQ001-1的制备
Step 1: Preparation of LQ001-1
物料配比：
Material ratio:
操作过程：Operation process:
反应瓶中加入8-溴辛酸、十七烷-9-醇、DCC、DMAP和DCM，室温下搅拌反应12h，TLC(EA:PE＝20:1)显示反应完全。 Add 8-bromooctanoic acid, heptadecan-9-ol, DCC, DMAP and DCM to the reaction bottle, and stir for 12 hours at room temperature. TLC (EA:PE=20:1) shows that the reaction is complete.
后处理：Post-processing:
反应液用硅藻土过滤后旋干，柱层析纯化后得40g无色油状物，收率为85％。The reaction solution was filtered through diatomaceous earth and spun to dryness. After purification by column chromatography, 40 g of colorless oil was obtained with a yield of 85%.
1HNMR(500MHz，CDCl3)δ：4.87(p，1H)，3.40(t，2H)，2.28(t，2H)，1.85(p，2H)，1.63(p，2H)，1.54-1.47(m，4H)，1.43(dt，2H)，1.33(dt，4H)，1.27(d，24H)，0.88(t，6H)。 1 HNMR (500MHz, CDCl 3 ) δ: 4.87 (p, 1H), 3.40 (t, 2H), 2.28 (t, 2H), 1.85 (p, 2H), 1.63 (p, 2H), 1.54-1.47 (m , 4H), 1.43 (dt, 2H), 1.33 (dt, 4H), 1.27 (d, 24H), 0.88 (t, 6H).
步骤2：LQ001-2的制备
Step 2: Preparation of LQ001-2
物料配比：
Material ratio:
操作过程：Operation process:
反应瓶中加入LQ001-1、乙醇胺和乙腈，加热至30℃搅拌反应12h。TLC(EA:PE＝20:1)显示反应完全。Add LQ001-1, ethanolamine and acetonitrile to the reaction bottle, heat to 30°C and stir for 12 hours. TLC (EA:PE=20:1) showed that the reaction was complete.
后处理：Post-processing:
反应液旋干后用500mL的乙酸乙酯稀释，用500mL水洗2次，取有机相用无水硫酸钠干燥后旋干，柱层析纯化后得到25g无色油状物，收率为87％。The reaction solution was spin-dried, diluted with 500 mL of ethyl acetate, and washed twice with 500 mL of water. The organic phase was dried over anhydrous sodium sulfate and then spin-dried. After purification by column chromatography, 25 g of colorless oil was obtained with a yield of 87%.
1HNMR(500MHz，CDCl3)δ：4.86(p，1H)，3.65-3.61(m，2H)，2.80-2.75(m，2H)，2.61(t，2H)，2.27(t，2H)，1.62(p，3H)，1.48(dt，7H)，1.32(s，7H)，1.27(d，22H)，0.87(t，6H)。 1 HNMR (500MHz, CDCl 3 ) δ: 4.86 (p, 1H), 3.65-3.61 (m, 2H), 2.80-2.75 (m, 2H), 2.61 (t, 2H), 2.27 (t, 2H), 1.62 (p, 3H), 1.48 (dt, 7H), 1.32 (s, 7H), 1.27 (d, 22H), 0.87 (t, 6H).
步骤3：LQ001-3的制备
Step 3: Preparation of LQ001-3
物料配比：

Material ratio:

操作过程：Operation process:
反应瓶中加入6-溴己酸、3,7-二甲基-1-辛醇、DCC、DMAP和DCM，室温下搅拌反应12h。TLC(EA:PE＝20:1)显示反应完全。Add 6-bromohexanoic acid, 3,7-dimethyl-1-octanol, DCC, DMAP and DCM to the reaction bottle, and stir for 12 hours at room temperature. TLC (EA:PE=20:1) showed that the reaction was complete.
后处理：Post-processing:
反应液用硅藻土过滤后旋干，柱层析纯化后得2.8g无色油状物，收率为83.6％。The reaction solution was filtered through diatomaceous earth and spun to dryness. After purification by column chromatography, 2.8 g of colorless oil was obtained, with a yield of 83.6%.
1HNMR(400MHz，CDCl3)δ：4.16-4.05(m，2H)，3.41(t，2H)，2.31(t，2H)，1.96-1.84(m，4H)，1.73-1.60(m，4H)，1.53-1.42(m，4H)，1.35-1.23(m，6H)，1.17-1.12(m，2H)，0.88(dd，9H)。 1 HNMR (400MHz, CDCl 3 ) δ: 4.16-4.05 (m, 2H), 3.41 (t, 2H), 2.31 (t, 2H), 1.96-1.84 (m, 4H), 1.73-1.60 (m, 4H) , 1.53-1.42(m, 4H), 1.35-1.23(m, 6H), 1.17-1.12(m, 2H), 0.88(dd, 9H).
步骤4：LQ001的制备
Step 4: Preparation of LQ001
物料配比：
Material ratio:
操作过程：Operation process:
反应瓶中加入LQ001-2、LQ001-3、K2CO3、KI、甲基叔丁基醚和乙腈，加热至80℃搅拌反应12h。TLC(DCM:MeOH＝10:1)显示反应完全。Add LQ001-2, LQ001-3, K 2 CO 3 , KI, methyl tert-butyl ether and acetonitrile to the reaction bottle, heat to 80°C and stir for 12 hours. TLC (DCM:MeOH=10:1) showed that the reaction was complete.
后处理：Post-processing:
反应液过滤后旋干，柱层析纯化后得到2g无色油状物，收率为77％。The reaction solution was filtered and spun to dryness. After purification by column chromatography, 2 g of colorless oil was obtained with a yield of 77%.
1HNMR(500MHz，CDCl3)δ：4.86(p，1H)，4.15-4.04(m，2H)，3.64(t，2H)，2.71(t，2H)，2.59(q，4H)，2.29(dt，4H)，1.64(tt，5H)，1.53(dq，10H)，1.42(dq，2H)，1.28(d，35H)，1.16-1.09(m，3H)，0.88(dd，15H)。 1 HNMR (500MHz, CDCl 3 ) δ: 4.86 (p, 1H), 4.15-4.04 (m, 2H), 3.64 (t, 2H), 2.71 (t, 2H), 2.59 (q, 4H), 2.29 (dt , 4H), 1.64 (tt, 5H), 1.53 (dq, 10H), 1.42 (dq, 2H), 1.28 (d, 35H), 1.16-1.09 (m, 3H), 0.88 (dd, 15H).
LCMS：Rt：4.991min；MS m/z(ESI)：697.1[M+H]+。LCMS: Rt: 4.991min; MS m/z (ESI): 697.1[M+H] + .
实施例2制备化合物LQ002Example 2 Preparation of Compound LQ002
步骤1：LQ002-1的制备
Step 1: Preparation of LQ002-1
物料配比：
Material ratio:
操作过程：Operation process:
反应瓶中加入8-溴辛酸、3,7-二甲基-1-辛醇、DCC、DMAP和DCM，室温下搅拌反应12h，TLC(EA:PE＝20:1)显示反应完全。Add 8-bromooctanoic acid, 3,7-dimethyl-1-octanol, DCC, DMAP and DCM to the reaction bottle, and stir for 12 hours at room temperature. TLC (EA:PE=20:1) shows that the reaction is complete.
后处理：Post-processing:
反应液用硅藻土过滤后旋干，柱层析纯化后得2g无色油状物，收率为55％。The reaction solution was filtered through diatomaceous earth and spun to dryness. After purification by column chromatography, 2 g of colorless oil was obtained with a yield of 55%.
1HNMR(500MHz，CDCl3)δ：4.17-4.03(m，2H)，3.40(t，2H)，2.29(t，2H)，1.85(dt，2H)，1.69-1.58(m，3H)，1.51(dt，2H)，1.42(dq，3H)，1.33(dt，4H)，1.30-1.19(m，3H)，1.17--1.07(m，3H)，0.88(dd，9H)。 1 HNMR (500MHz, CDCl 3 ) δ: 4.17-4.03 (m, 2H), 3.40 (t, 2H), 2.29 (t, 2H), 1.85 (dt, 2H), 1.69-1.58 (m, 3H), 1.51 (dt, 2H), 1.42 (dq, 3H), 1.33 (dt, 4H), 1.30-1.19 (m, 3H), 1.17--1.07 (m, 3H), 0.88 (dd, 9H).
步骤2：LQ002-2的制备
Step 2: Preparation of LQ002-2
物料配比：
Material ratio:
操作过程：Operation process:
反应瓶中加入LQ002-1、乙醇胺和乙腈，室温下搅拌反应12h。TLC(EA:PE＝20:1)显示反应完全。Add LQ002-1, ethanolamine and acetonitrile to the reaction bottle, and stir for 12 hours at room temperature. TLC (EA:PE=20:1) showed that the reaction was complete.
后处理：Post-processing:
反应液旋干后用100mL的乙酸乙酯稀释，用100mL水洗2次，取有机相用无水硫酸钠干燥后旋干，柱层析纯化后得到750mg无色油状物，收率为57.8％。The reaction solution was spin-dried, diluted with 100 mL of ethyl acetate, and washed twice with 100 mL of water. The organic phase was dried over anhydrous sodium sulfate and then spin-dried. After purification by column chromatography, 750 mg of colorless oil was obtained, with a yield of 57.8%.
TLC情况：DCM:MeOH＝10:1，Rf＝0.4。TLC case: DCM:MeOH=10:1, Rf=0.4.
步骤3：LQ002的制备
Step 3: Preparation of LQ002
物料配比：
Material ratio:
操作过程：Operation process:
反应瓶中加入LQ002-2、LQ001-3、K2CO3、KI、甲基叔丁基醚和乙腈，加热至80℃搅拌反应12h。TLC(DCM:MeOH＝10:1)显示反应完全。Add LQ002-2, LQ001-3, K 2 CO 3 , KI, methyl tert-butyl ether and acetonitrile to the reaction bottle, heat to 80°C and stir for 12 hours. TLC (DCM:MeOH=10:1) showed that the reaction was complete.
后处理：Post-processing:
反应液过滤后旋干，柱层析纯化后得到1g无色油状物，收率为76％。The reaction solution was filtered and spun to dryness. After purification by column chromatography, 1 g of colorless oil was obtained with a yield of 76%.
1HNMR(400MHz，CDCl3)δ：4.09(q，4H)，3.58(t，2H)，2.64(t，2H)，2.56-2.48(m，4H)，2.32-2.25(m，4H)，1.69-1.57(m，6H)，1.51(dt，7H)，1.42(dt，4H)，1.34-1.20(m，14H)，1.18-1.08(m，6H)，0.87(dd，18H)。 1 HNMR (400MHz, CDCl 3 ) δ: 4.09 (q, 4H), 3.58 (t, 2H), 2.64 (t, 2H), 2.56-2.48 (m, 4H), 2.32-2.25 (m, 4H), 1.69 -1.57 (m, 6H), 1.51 (dt, 7H), 1.42 (dt, 4H), 1.34-1.20 (m, 14H), 1.18-1.08 (m, 6H), 0.87 (dd, 18H).
LCMS：Rt：2.429min；MS m/z(ESI)：598.9[M+H]+。LCMS: Rt: 2.429min; MS m/z (ESI): 598.9[M+H] + .
实施例3制备化合物LQ004Example 3 Preparation of Compound LQ004
步骤1：LQ004-1的制备
Step 1: Preparation of LQ004-1
物料配比：
Material ratio:
操作过程：Operation process:
反应瓶中加入6-溴己酸、2-环己基乙醇、DCC、DMAP和DCM，室温下搅拌反应12h。TLC(EA:PE＝20:1)显示反应完全。Add 6-bromohexanoic acid, 2-cyclohexylethanol, DCC, DMAP and DCM to the reaction bottle, and stir for 12 hours at room temperature. TLC (EA:PE=20:1) showed that the reaction was complete.
后处理：Post-processing:
反应液用硅藻土过滤后旋干，柱层析纯化后得2g无色油状物。The reaction solution was filtered through diatomaceous earth and spun to dryness. After purification by column chromatography, 2 g of colorless oil was obtained.
TLC情况：EA:PE＝20:1，Rf＝0.6。TLC situation: EA:PE=20:1, Rf=0.6.
步骤2：LQ004的制备
Step 2: Preparation of LQ004
物料配比：
Material ratio:
操作过程：Operation process:
反应瓶中加入LQ001-2、LQ004-1、K2CO3、KI、甲基叔丁基醚和乙腈，加热至80℃搅拌反应12h。TLC(DCM:MeOH＝10:1)显示反应完全。Add LQ001-2, LQ004-1, K 2 CO 3 , KI, methyl tert-butyl ether and acetonitrile into the reaction bottle, heat to 80°C and stir for 12 hours. TLC (DCM:MeOH=10:1) showed that the reaction was complete.
后处理：Post-processing:
反应液过滤后旋干，柱层析纯化后得到2g无色油状物，收率为80％。The reaction solution was filtered and spun to dryness. After purification by column chromatography, 2 g of colorless oil was obtained with a yield of 80%.
1HNMR(500MHz，CDCl3)δ：4.86(p，1H)，4.10(t，2H)，3.64-3.58(m，2H)，2.67(s，2H)， 2.55(s，4H)，2.29(dt，4H)，1.65(dddd，11H)，1.51(q，11H)，1.32-1.23(m，33H)，0.88(t，9H)。 1 HNMR (500MHz, CDCl 3 ) δ: 4.86 (p, 1H), 4.10 (t, 2H), 3.64-3.58 (m, 2H), 2.67 (s, 2H), 2.55(s, 4H), 2.29(dt, 4H), 1.65(dddd, 11H), 1.51(q, 11H), 1.32-1.23(m, 33H), 0.88(t, 9H).
LCMS：Rt：5.913min；MS m/z(ESI)：566.9[M+H]+。LCMS: Rt: 5.913min; MS m/z (ESI): 566.9[M+H] + .
实施例4制备化合物LQ005Example 4 Preparation of Compound LQ005
步骤1：LQ005-1的制备
Step 1: Preparation of LQ005-1
物料配比：
Material ratio:
操作过程：Operation process:
反应瓶中加入8-溴辛酸、2-环己基乙醇、DCC、DMAP和DCM，室温下搅拌反应12h，TLC(EA:PE＝20:1)显示反应完全。Add 8-bromooctanoic acid, 2-cyclohexylethanol, DCC, DMAP and DCM to the reaction bottle, and stir for 12 hours at room temperature. TLC (EA:PE=20:1) shows that the reaction is complete.
后处理：Post-processing:
反应液用硅藻土过滤后旋干，柱层析纯化后得2g无色油状物，收率为55％。The reaction solution was filtered through diatomaceous earth and spun to dryness. After purification by column chromatography, 2 g of colorless oil was obtained with a yield of 55%.
TLC情况：EA:PE＝20:1，Rf＝0.6。TLC situation: EA:PE=20:1, Rf=0.6.
步骤2：LQ005-2的制备
Step 2: Preparation of LQ005-2
物料配比：

Material ratio:

操作过程：Operation process:
反应瓶中加入LQ005-1、乙醇胺和乙腈，室温下搅拌反应12h。TLC(EA:PE＝20:1)显示反应完全。Add LQ005-1, ethanolamine and acetonitrile to the reaction bottle, and stir for 12 hours at room temperature. TLC (EA:PE=20:1) showed that the reaction was complete.
后处理：Post-processing:
反应液旋干后用100ml的乙酸乙酯稀释，用100ml水洗2次，取有机相用无水硫酸钠干燥后旋干，柱层析纯化后得到750mg无色油状物，收率为57.8％。The reaction solution was spin-dried, diluted with 100 ml of ethyl acetate, and washed twice with 100 ml of water. The organic phase was dried over anhydrous sodium sulfate and then spun-dried. After purification by column chromatography, 750 mg of colorless oil was obtained, with a yield of 57.8%.
TLC情况：DCM:MeOH＝10:1，Rf＝0.4。TLC case: DCM:MeOH=10:1, Rf=0.4.
步骤3：LQ005的制备
Step 3: Preparation of LQ005
物料配比：
Material ratio:
操作过程：Operation process:
反应瓶中加入LQ005-2、LQ004-1、K2CO3、KI、甲基叔丁基醚和乙腈，加热至80℃搅拌反应12h。TLC(DCM:MeOH＝10:1)显示反应完全，产物Rf＝0.4。Add LQ005-2, LQ004-1, K 2 CO 3 , KI, methyl tert-butyl ether and acetonitrile to the reaction bottle, heat to 80°C and stir for 12 hours. TLC (DCM:MeOH=10:1) showed that the reaction was complete and the product Rf=0.4.
后处理：Post-processing:
反应液过滤后旋干，柱层析纯化后得到1g无色油状物，收率为76％。The reaction solution was filtered and spun to dryness. After purification by column chromatography, 1 g of colorless oil was obtained with a yield of 76%.
1HNMR(400MHz，CDCl3)δ：4.09(t，4H)，3.67-3.60(m，2H)，2.75-2.68(m，2H)，2.65-2.54(m，4H)，2.29(q，4H)，1.73-1.58(m，15H)，1.51(q，6H)，1.40-1.08(m，18H)，0.97-0.86(m，4H)。 1 HNMR (400MHz, CDCl 3 ) δ: 4.09 (t, 4H), 3.67-3.60 (m, 2H), 2.75-2.68 (m, 2H), 2.65-2.54 (m, 4H), 2.29 (q, 4H) , 1.73-1.58(m, 15H), 1.51(q, 6H), 1.40-1.08(m, 18H), 0.97-0.86(m, 4H).
LCMS：Rt：0.086min；MS m/z(ESI)：538.7[M+H]+。 LCMS: Rt: 0.086min; MS m/z (ESI): 538.7[M+H] + .
实施例5制备化合物LQ007Example 5 Preparation of Compound LQ007
步骤1：LQ007-1的制备
Step 1: Preparation of LQ007-1
物料配比：
Material ratio:
操作过程：Operation process:
反应瓶中加入6-溴己酸、异癸醇、DCC、DMAP和DCM，室温下搅拌反应12h。TLC(EA:PE＝20:1)显示反应完全。Add 6-bromohexanoic acid, isodecanol, DCC, DMAP and DCM to the reaction bottle, and stir for 12 hours at room temperature. TLC (EA:PE=20:1) showed that the reaction was complete.
后处理：Post-processing:
反应液用硅藻土过滤后旋干，柱层析纯化后得2g无色油状物。The reaction solution was filtered through diatomaceous earth and spun to dryness. After purification by column chromatography, 2 g of colorless oil was obtained.
TLC情况：EA:PE＝20:1,产物Rf＝0.6。TLC situation: EA:PE=20:1, product Rf=0.6.
步骤2：LQ007的制备
Step 2: Preparation of LQ007
物料配比：
Material ratio:
操作过程： Operation process:
反应瓶中加入LQ001-2、LQ007-1、K2CO3、KI、甲基叔丁基醚和乙腈，加热至80℃搅拌反应12h。TLC(DCM:MeOH＝10:1)显示反应完全。Add LQ001-2, LQ007-1, K 2 CO 3 , KI, methyl tert-butyl ether and acetonitrile to the reaction bottle, heat to 80°C and stir for 12 hours. TLC (DCM:MeOH=10:1) showed that the reaction was complete.
后处理：Post-processing:
反应液过滤后旋干，柱层析纯化后得到2g无色油状物，收率为80％。The reaction solution was filtered and spun to dryness. After purification by column chromatography, 2 g of colorless oil was obtained with a yield of 80%.
1HNMR(500MHz，CDCl3)δ：4.86(p，1H)，4.14-4.02(m，2H)，3.56(t，2H)，2.61(t，2H)，2.48(q，4H)，2.32-2.25(m，4H)，1.68-1.58(m，6H)，1.56-1.40(m，10H)，1.36-1.21(m，38H)，1.19-1.03(m，4H)，0.87(t，12H)。 1 HNMR (500MHz, CDCl 3 ) δ: 4.86 (p, 1H), 4.14-4.02 (m, 2H), 3.56 (t, 2H), 2.61 (t, 2H), 2.48 (q, 4H), 2.32-2.25 (m, 4H), 1.68-1.58 (m, 6H), 1.56-1.40 (m, 10H), 1.36-1.21 (m, 38H), 1.19-1.03 (m, 4H), 0.87 (t, 12H).
LCMS：Rt：3.83min；MS m/z(ESI)：696.7[M+H]+。LCMS: Rt: 3.83min; MS m/z (ESI): 696.7[M+H] + .
实施例6制备化合物LQ008Example 6 Preparation of Compound LQ008
步骤1：LQ008-1的制备
Step 1: Preparation of LQ008-1
物料配比：
Material ratio:
操作过程：Operation process:
反应瓶中加入8-溴辛酸、异癸醇、DCC、DMAP和DCM，室温下搅拌反应12h，TLC(EA:PE＝20:1)显示反应完全。Add 8-bromooctanoic acid, isodecyl alcohol, DCC, DMAP and DCM to the reaction bottle, and stir for 12 hours at room temperature. TLC (EA:PE=20:1) shows that the reaction is complete.
后处理：Post-processing:
反应液用硅藻土过滤后旋干，柱层析纯化后得2g无色油状物，收率为55％。The reaction solution was filtered through diatomaceous earth and spun to dryness. After purification by column chromatography, 2 g of colorless oil was obtained with a yield of 55%.
步骤2：LQ008-2的制备
Step 2: Preparation of LQ008-2
物料配比：
Material ratio:
操作过程：Operation process:
反应瓶中加入LQ008-1、乙醇胺和乙腈，室温下搅拌反应12h。TLC(EA:PE＝20:1)显示反应完全。Add LQ008-1, ethanolamine and acetonitrile to the reaction bottle, and stir for 12 hours at room temperature. TLC (EA:PE=20:1) showed that the reaction was complete.
后处理：Post-processing:
反应液旋干后用100ml的乙酸乙酯稀释，用100ml水洗2次，取有机相用无水硫酸钠干燥后旋干，柱层析纯化后得到750mg无色油状物，收率57.8％。The reaction solution was spin-dried, diluted with 100 ml of ethyl acetate, and washed twice with 100 ml of water. The organic phase was dried over anhydrous sodium sulfate and then spun-dried. After purification by column chromatography, 750 mg of colorless oil was obtained, with a yield of 57.8%.
1HNMR(500MHz，CDCl3)δ：4.16-3.99(m，2H)，3.69-3.60(m，2H)，2.85-2.73(m，2H)，2.63(t，2H)，2.28(td，2H)，2.06(s，4H)，1.66-1.57(m，3H)，1.49(q，2H)，1.35-1.24(m，2H)，1.16-1.05(m，2H)，0.85(tq，8H)。 1 HNMR (500MHz, CDCl 3 ) δ: 4.16-3.99 (m, 2H), 3.69-3.60 (m, 2H), 2.85-2.73 (m, 2H), 2.63 (t, 2H), 2.28 (td, 2H) , 2.06 (s, 4H), 1.66-1.57 (m, 3H), 1.49 (q, 2H), 1.35-1.24 (m, 2H), 1.16-1.05 (m, 2H), 0.85 (tq, 8H).
步骤3：LQ008的制备
Step 3: Preparation of LQ008
物料配比：
Material ratio:
操作过程：Operation process:
反应瓶中加入LQ008-2、LQ007-1、K2CO3、KI、甲基叔丁基醚和乙腈，加热至80℃搅拌反应12h。TLC(DCM:MeOH＝10:1)显示反应完全，产物Rf＝0.4。Add LQ008-2, LQ007-1, K 2 CO 3 , KI, methyl tert-butyl ether and acetonitrile to the reaction bottle, heat to 80°C and stir for 12 hours. TLC (DCM:MeOH=10:1) showed that the reaction was complete and the product Rf=0.4.
后处理：Post-processing:
反应液过滤后旋干，柱层析纯化后得到1g无色油状物，收率为76％。The reaction solution was filtered and spun to dryness. After purification by column chromatography, 1 g of colorless oil was obtained with a yield of 76%.
1HNMR(500MHz，CDCl3)δ：4.17-4.01(m，3H)，3.60(t，2H)，2.66(s，2H)，2.54(d，4H)， 2.29(qd，4H)，1.75-1.44(m，13H)，1.30(d，18H)，1.19-0.97(m，5H)，0.94-0.71(m，18H)。 1 HNMR (500MHz, CDCl 3 ) δ: 4.17-4.01 (m, 3H), 3.60 (t, 2H), 2.66 (s, 2H), 2.54 (d, 4H), 2.29 (qd, 4H), 1.75-1.44 (m, 13H), 1.30 (d, 18H), 1.19-0.97 (m, 5H), 0.94-0.71 (m, 18H).
LCMS：Rt：4.452min；MS m/z(ESI)：598.9[M+H]+。LCMS: Rt: 4.452min; MS m/z (ESI): 598.9[M+H] + .
实施例7制备化合物LQ010Example 7 Preparation of Compound LQ010
步骤1：LQ010-1的制备
Step 1: Preparation of LQ010-1
物料配比：
Material ratio:
操作过程：Operation process:
反应瓶中加入6-溴己酸、3,5,5-三甲基-1-己醇、DCC、DMAP和DCM，室温下搅拌反应12h。TLC(EA:PE＝20:1)显示反应完全。Add 6-bromohexanoic acid, 3,5,5-trimethyl-1-hexanol, DCC, DMAP and DCM to the reaction bottle, and stir for 12 hours at room temperature. TLC (EA:PE=20:1) showed that the reaction was complete.
后处理：Post-processing:
反应液用硅藻土过滤后旋干，柱层析纯化后得2g无色油状物。The reaction solution was filtered through diatomaceous earth and spun to dryness. After purification by column chromatography, 2 g of colorless oil was obtained.
TLC情况：EA:PE＝20:1，Rf＝0.6。TLC situation: EA:PE=20:1, Rf=0.6.
步骤2：LQ010的制备
Step 2: Preparation of LQ010
物料配比：
Material ratio:
操作过程：Operation process:
反应瓶中加入LQ001-2、LQ010-1、K2CO3、KI、甲基叔丁基醚和乙腈，加热至80℃搅拌反应12h。TLC(DCM:MeOH＝10:1)显示反应完全，产物Rf＝0.4。Add LQ001-2, LQ010-1, K 2 CO 3 , KI, methyl tert-butyl ether and acetonitrile to the reaction bottle, heat to 80°C and stir for 12 hours. TLC (DCM:MeOH=10:1) showed that the reaction was complete and the product Rf=0.4.
后处理：Post-processing:
反应液过滤后旋干，柱层析纯化后得到2g无色油状物，收率为80％。The reaction solution was filtered and spun to dryness. After purification by column chromatography, 2 g of colorless oil was obtained with a yield of 80%.
1HNMR(500MHz，CDCl3)δ：4.85(p，1H)，4.12-4.03(m，2H)，3.72(t，2H)，2.84-2.79(m，2H)，2.74-2.67(m，4H)，2.29(dt，4H)，1.67-1.57(m，10H)，1.47(dd，6H)，1.38-1.23(m，33H)，1.07(dd，1H)，0.94(d，3H)，0.88(t，15H)。 1 HNMR (500MHz, CDCl 3 ) δ: 4.85 (p, 1H), 4.12-4.03 (m, 2H), 3.72 (t, 2H), 2.84-2.79 (m, 2H), 2.74-2.67 (m, 4H) , 2.29(dt, 4H), 1.67-1.57(m, 10H), 1.47(dd, 6H), 1.38-1.23(m, 33H), 1.07(dd, 1H), 0.94(d, 3H), 0.88(t ,15H).
LCMS：Rt：3.83min；MS m/z(ESI)：682.7[M+H]+。LCMS: Rt: 3.83min; MS m/z (ESI): 682.7[M+H] + .
实施例8制备化合物LQ011Example 8 Preparation of Compound LQ011
步骤1：LQ011-1的制备
Step 1: Preparation of LQ011-1
物料配比：
Material ratio:
操作过程：Operation process:
反应瓶中加入8-溴辛酸、3,5,5-三甲基-1-己醇、DCC、DMAP和DCM，室温下搅拌反应12h，TLC(EA:PE＝20:1)显示反应完全。Add 8-bromooctanoic acid, 3,5,5-trimethyl-1-hexanol, DCC, DMAP and DCM to the reaction bottle, and stir for 12 hours at room temperature. TLC (EA:PE=20:1) shows that the reaction is complete.
后处理：Post-processing:
反应液用硅藻土过滤后旋干，柱层析纯化后得2g无色油状。The reaction solution was filtered through diatomaceous earth and spun to dryness. After purification by column chromatography, 2 g of colorless oil was obtained.
TLC情况：EA:PE＝20:1，产物Rf＝0.6。TLC situation: EA:PE=20:1, product Rf=0.6.
步骤2：LQ011-2的制备
Step 2: Preparation of LQ011-2
物料配比：
Material ratio:
操作过程：Operation process:
反应瓶中加入LQ011-1、乙醇胺和乙腈，室温下搅拌反应12h。TLC(EA:PE＝20:1)显示反应完全。Add LQ011-1, ethanolamine and acetonitrile to the reaction bottle, and stir for 12 hours at room temperature. TLC (EA:PE=20:1) showed that the reaction was complete.
后处理：Post-processing:
反应液旋干后用100mL的乙酸乙酯稀释，用100ml水洗2次，取有机相用无水硫酸钠干燥后旋干，柱层析纯化后得到750mg无色油状物，收率为57.8％。The reaction solution was spin-dried, diluted with 100 mL of ethyl acetate, and washed twice with 100 ml of water. The organic phase was dried over anhydrous sodium sulfate and then spin-dried. After purification by column chromatography, 750 mg of colorless oil was obtained, with a yield of 57.8%.
1HNMR(500MHz，CDCl3)δ：4.08(t，2H)，3.64(t，2H)，2.78(t，2H)，2.62(t，2H)，2.28(t，4H)，1.66-1.56(m，4H)，1.47(dt，3H)，1.31(s，6H)，1.22(dd，1H)，1.07(dd，1H)，0.94(d，3H)，0.88(s，9H)。 1 HNMR (500MHz, CDCl 3 ) δ: 4.08 (t, 2H), 3.64 (t, 2H), 2.78 (t, 2H), 2.62 (t, 2H), 2.28 (t, 4H), 1.66-1.56 (m , 4H), 1.47 (dt, 3H), 1.31 (s, 6H), 1.22 (dd, 1H), 1.07 (dd, 1H), 0.94 (d, 3H), 0.88 (s, 9H).
步骤3：LQ011的制备
Step 3: Preparation of LQ011
物料配比：

Material ratio:

操作过程：Operation process:
反应瓶中加入LQ008-2、LQ007-1、K2CO3、KI、甲基叔丁基醚和乙腈，加热至80℃搅拌反应12h。TLC(DCM:MeOH＝10:1)显示反应完全，产物Rf＝0.4。Add LQ008-2, LQ007-1, K 2 CO 3 , KI, methyl tert-butyl ether and acetonitrile to the reaction bottle, heat to 80°C and stir for 12 hours. TLC (DCM:MeOH=10:1) showed that the reaction was complete and the product Rf=0.4.
后处理：Post-processing:
反应液过滤后旋干，柱层析纯化后得到1g无色油状物，收率为76％。The reaction solution was filtered and spun to dryness. After purification by column chromatography, 1 g of colorless oil was obtained with a yield of 76%.
1HNMR(500MHz，CDCl3)δ：4.07(t，4H)，3.60(t，2H)，2.67(t，2H)，2.54(q，4H)，2.28(q，4H)，1.61(ddd，8H)，1.54-1.43(m，6H)，1.30(q，8H)，1.22(dd，2H)，1.07(dd，2H)，0.93(d，6H)，0.88(t，18H)。 1 HNMR (500MHz, CDCl 3 ) δ: 4.07 (t, 4H), 3.60 (t, 2H), 2.67 (t, 2H), 2.54 (q, 4H), 2.28 (q, 4H), 1.61 (ddd, 8H ), 1.54-1.43(m, 6H), 1.30(q, 8H), 1.22(dd, 2H), 1.07(dd, 2H), 0.93(d, 6H), 0.88(t, 18H).
LCMS：Rt：1.609min；MS m/z(ESI)：570.5[M]+。LCMS: Rt: 1.609min; MS m/z (ESI): 570.5[M] + .
实施例9制备化合物LQ013Example 9 Preparation of Compound LQ013
步骤1：LQ013-1的制备
Step 1: Preparation of LQ013-1
物料配比：
Material ratio:
操作过程：Operation process:
反应瓶中加入6-溴己酸、2-乙基己醇、DCC、DMAP和DCM，室温下搅拌反应12h。TLC(EA:PE＝20:1)显示反应完全。Add 6-bromohexanoic acid, 2-ethylhexanol, DCC, DMAP and DCM to the reaction bottle, and stir for 12 hours at room temperature. TLC (EA:PE=20:1) showed that the reaction was complete.
后处理：Post-processing:
反应液用硅藻土过滤后旋干，柱层析纯化后得2g无色油状物。The reaction solution was filtered through diatomaceous earth and spun to dryness. After purification by column chromatography, 2 g of colorless oil was obtained.
TLC情况：EA:PE＝20:1，产物Rf＝0.6。TLC situation: EA:PE=20:1, product Rf=0.6.
步骤2：LQ013的制备
Step 2: Preparation of LQ013
物料配比：
Material ratio:
操作过程：Operation process:
反应瓶中加入LQ001-2、LQ013-1、K2CO3、KI、甲基叔丁基醚和乙腈，加热至80℃搅拌反应12h。TLC(DCM:MeOH＝10:1)显示反应完全，产物Rf＝0.4。Add LQ001-2, LQ013-1, K 2 CO 3 , KI, methyl tert-butyl ether and acetonitrile to the reaction bottle, heat to 80°C and stir for 12 hours. TLC (DCM:MeOH=10:1) showed that the reaction was complete and the product Rf=0.4.
后处理：Post-processing:
反应液过滤后旋干，柱层析纯化后得到2g无色油状物，收率为80％。The reaction solution was filtered and spun to dryness. After purification by column chromatography, 2 g of colorless oil was obtained with a yield of 80%.
1HNMR(500MHz，CDCl3)δ：4.92-4.85(m，1H)，4.01(m，2H)，3.72(s，2H)，2.81(s，2H)，2.70(s，4H)，2.37-2.28(m，4H)，1.73-1.49(m，15H)，1.38-1.26(m，39H)，0.91(td，12H)。 1 HNMR (500MHz, CDCl 3 ) δ: 4.92-4.85 (m, 1H), 4.01 (m, 2H), 3.72 (s, 2H), 2.81 (s, 2H), 2.70 (s, 4H), 2.37-2.28 (m, 4H), 1.73-1.49 (m, 15H), 1.38-1.26 (m, 39H), 0.91 (td, 12H).
实施例10制备化合物LQ014Example 10 Preparation of Compound LQ014
步骤1：LQ014-1的制备
Step 1: Preparation of LQ014-1
物料配比：

Material ratio:

操作过程：Operation process:
反应瓶中加入8-溴辛酸、2-乙基己醇、DCC、DMAP和DCM，室温下搅拌反应12h，TLC(EA:PE＝20:1)显示反应完全。Add 8-bromooctanoic acid, 2-ethylhexanol, DCC, DMAP and DCM to the reaction bottle, and stir for 12 hours at room temperature. TLC (EA:PE=20:1) shows that the reaction is complete.
后处理：Post-processing:
反应液用硅藻土过滤后旋干，柱层析纯化后得2g无色油状物，收率为55％。The reaction solution was filtered through diatomaceous earth and spun to dryness. After purification by column chromatography, 2 g of colorless oil was obtained with a yield of 55%.
步骤2：LQ014-2的制备
Step 2: Preparation of LQ014-2
物料配比：
Material ratio:
操作过程：Operation process:
反应瓶中加入LQ014-1、乙醇胺和乙腈，室温下搅拌反应12h。TLC(EA:PE＝20:1)显示反应完全。Add LQ014-1, ethanolamine and acetonitrile to the reaction bottle, and stir for 12 hours at room temperature. TLC (EA:PE=20:1) showed that the reaction was complete.
后处理：Post-processing:
反应液旋干后用100mL的乙酸乙酯稀释，用100mL水洗2次，取有机相用无水硫酸钠干燥后旋干，柱层析纯化后得到750mg无色油状物，收率为55％。The reaction solution was spin-dried, diluted with 100 mL of ethyl acetate, and washed twice with 100 mL of water. The organic phase was dried over anhydrous sodium sulfate and then spin-dried. After purification by column chromatography, 750 mg of colorless oil was obtained, with a yield of 55%.
TLC情况：DCM:MeOH＝10:1，产物Rf＝0.2。TLC situation: DCM:MeOH=10:1, product Rf=0.2.
步骤3：LQ014的制备
Step 3: Preparation of LQ014
物料配比：
Material ratio:
操作过程：Operation process:
反应瓶中加入LQ014-2、LQ013-1、K2CO3、KI、甲基叔丁基醚和乙腈，加热至80℃搅拌反应12h。TLC(DCM:MeOH＝10:1)显示反应完全，产物Rf＝0.4。Add LQ014-2, LQ013-1, K 2 CO 3 , KI, methyl tert-butyl ether and acetonitrile to the reaction bottle, heat to 80°C and stir for 12 hours. TLC (DCM:MeOH=10:1) showed that the reaction was complete and the product Rf=0.4.
后处理：Post-processing:
反应液过滤后旋干，柱层析纯化后得到1g无色油状物，收率为73％。The reaction solution was filtered and spun to dryness. After purification by column chromatography, 1 g of colorless oil was obtained with a yield of 73%.
1HNMR(500MHz，CDCl3)δ：4.03-3.93(m，4H)，3.65-3.55(m，2H)，2.66(s，2H)，2.53(s，4H)，2.30(td，4H)，1.69-1.47(m，11H)，1.39-1.26(m，24H)，0.94-0.85(m，12H)。 1 HNMR (500MHz, CDCl 3 ) δ: 4.03-3.93 (m, 4H), 3.65-3.55 (m, 2H), 2.66 (s, 2H), 2.53 (s, 4H), 2.30 (td, 4H), 1.69 -1.47 (m, 11H), 1.39-1.26 (m, 24H), 0.94-0.85 (m, 12H).
实施例11制备化合物LQ016Example 11 Preparation of Compound LQ016
步骤1：LQ016-1的制备
Step 1: Preparation of LQ016-1
物料配比：
Material ratio:
操作过程： Operation process:
反应瓶中加入6-溴己酸、1,2-环氧十二烷、三氯化铁和吡啶，室温下搅拌反应12h。TLC(EA:PE＝4:1)显示反应完全。Add 6-bromohexanoic acid, 1,2-epoxydodecane, ferric chloride and pyridine to the reaction bottle, and stir for 12 hours at room temperature. TLC (EA:PE=4:1) showed that the reaction was complete.
后处理：Post-processing:
反应液用柱层析纯化后得1.4g产物。The reaction solution was purified by column chromatography to obtain 1.4 g of product.
TLC情况：EA:PE＝4:1，产物Rf＝0.5。TLC situation: EA:PE=4:1, product Rf=0.5.
步骤2：LQ016的制备
Step 2: Preparation of LQ016
物料配比：
Material ratio:
操作过程：Operation process:
反应瓶中加入LQ001-2、LQ016-1、K2CO3、KI、甲基叔丁基醚和乙腈，加热至80℃搅拌反应12h。TLC(DCM:MeOH＝10:1)显示反应完全，产物Rf＝0.4。Add LQ001-2, LQ016-1, K 2 CO 3 , KI, methyl tert-butyl ether and acetonitrile to the reaction bottle, heat to 80°C and stir for 12 hours. TLC (DCM:MeOH=10:1) showed that the reaction was complete and the product Rf=0.4.
后处理：Post-processing:
反应液过滤后旋干，柱层析纯化后得到900g无色油状物。The reaction solution was filtered, spun to dryness, and purified by column chromatography to obtain 900 g of colorless oil.
1HNMR(500MHz，CDCl3)δ：4.86(p，1H)，4.15(dd，1H)，3.94(dd，1H)，3.87-3.80(m，1H)，3.65-3.58(m，2H)，2.67(s，2H)，2.56(s，4H)，2.36(td，2H)，2.28(t，2H)，1.71-1.43(m，16H)，1.27(d，46H)，0.87(t，9H)。 1 HNMR (500MHz, CDCl 3 ) δ: 4.86 (p, 1H), 4.15 (dd, 1H), 3.94 (dd, 1H), 3.87-3.80 (m, 1H), 3.65-3.58 (m, 2H), 2.67 (s, 2H), 2.56 (s, 4H), 2.36 (td, 2H), 2.28 (t, 2H), 1.71-1.43 (m, 16H), 1.27 (d, 46H), 0.87 (t, 9H).
LCMS：Rt：3.83min；MS m/z(ESI)：740.7[M]+。LCMS: Rt: 3.83min; MS m/z (ESI): 740.7[M] + .
实施例12制备化合物LQ017Example 12 Preparation of Compound LQ017
步骤1：LQ017-1的制备
Step 1: Preparation of LQ017-1
物料配比：
Material ratio:
操作过程：Operation process:
反应瓶中加入8-溴辛酸、1,2-环氧十二烷、三氯化铁和吡啶，室温下搅拌反应12h，TLC(EA:PE＝4:1)显示反应完全。Add 8-bromooctanoic acid, 1,2-epoxydodecane, ferric chloride and pyridine to the reaction bottle, and stir for 12 hours at room temperature. TLC (EA:PE=4:1) shows that the reaction is complete.
后处理：Post-processing:
反应液用硅藻土过滤后旋干，柱层析纯化后得1g无色油状物，收率为25％。The reaction solution was filtered through diatomaceous earth and spun to dryness. After purification by column chromatography, 1 g of colorless oil was obtained with a yield of 25%.
1HNMR(400MHz，CDCl3)δ：4.91(ddt，1H)，3.77-3.56(m，2H)，3.40(t，2H)，2.34(t，2H)，1.85(dt，3H)，1.68-1.52(m，6H)，1.43(dd，2H)，1.34(dt，5H)，1.30-1.23(m，13H)，0.88(t，3H)。 1 HNMR (400MHz, CDCl 3 ) δ: 4.91 (ddt, 1H), 3.77-3.56 (m, 2H), 3.40 (t, 2H), 2.34 (t, 2H), 1.85 (dt, 3H), 1.68-1.52 (m, 6H), 1.43 (dd, 2H), 1.34 (dt, 5H), 1.30-1.23 (m, 13H), 0.88 (t, 3H).
步骤2：LQ017-2的制备
Step 2: Preparation of LQ017-2
物料配比：
Material ratio:
操作过程：Operation process:
反应瓶中加入LQ017-1、乙醇胺和乙腈，室温下搅拌反应2d。TLC(EA:PE＝4:1)显示反应完全。Add LQ017-1, ethanolamine and acetonitrile to the reaction bottle, and stir for 2 days at room temperature. TLC (EA:PE=4:1) showed that the reaction was complete.
后处理： Post-processing:
反应液旋干后用100mL的乙酸乙酯稀释，用100mL水洗2次，取有机相用无水硫酸钠干燥后旋干，柱层析纯化后得到300mg无色油状物，收率为32.8％。The reaction solution was spin-dried, diluted with 100 mL of ethyl acetate, and washed twice with 100 mL of water. The organic phase was dried over anhydrous sodium sulfate and then spin-dried. After purification by column chromatography, 300 mg of colorless oil was obtained, with a yield of 32.8%.
1HNMR(400MHz，CDCl3)δ：4.15(dd，1H)，3.94(dd，1H)，3.87-3.79(m，1H)，3.71-3.66(m，2H)，2.85-2.79(m，2H)，2.66(t，2H)，2.35(t，2H)，1.70-1.40(m，8H)，1.29(d，22H)，0.88(t，3H)。 1 HNMR (400MHz, CDCl 3 ) δ: 4.15 (dd, 1H), 3.94 (dd, 1H), 3.87-3.79 (m, 1H), 3.71-3.66 (m, 2H), 2.85-2.79 (m, 2H) , 2.66(t, 2H), 2.35(t, 2H), 1.70-1.40(m, 8H), 1.29(d, 22H), 0.88(t, 3H).
步骤3：LQ017的制备
Step 3: Preparation of LQ017
物料配比：
Material ratio:
操作过程：Operation process:
反应瓶中加入LQ017-2、LQ016-1、K2CO3、KI、甲基叔丁基醚和乙腈，加热至80℃搅拌反应12h。TLC(DCM:MeOH＝10:1)显示反应完全，产物Rf＝0.4。Add LQ017-2, LQ016-1, K 2 CO 3 , KI, methyl tert-butyl ether and acetonitrile to the reaction bottle, heat to 80°C and stir for 12 hours. TLC (DCM:MeOH=10:1) showed that the reaction was complete and the product Rf=0.4.
后处理：Post-processing:
反应液过滤后旋干，柱层析纯化后得到160mg无色油状物，收率为76％。The reaction solution was filtered and spun to dryness. After purification by column chromatography, 160 mg of colorless oil was obtained, with a yield of 76%.
1HNMR(400MHz，CDCl3)δ：4.14(dd，2H)，4.07-4.02(m，2H)，3.96(ddd，2H)，3.89-3.81(m，2H)，3.25-3.19(m，2H)，3.17-3.09(m，4H)，2.38(dt，4H)，1.94-1.79(m，5H)，1.77-1.58(m，6H)，1.52-1.41(m，8H)，1.39-1.24(m，36H)，0.87(t，6H)。 1 HNMR (400MHz, CDCl 3 ) δ: 4.14 (dd, 2H), 4.07-4.02 (m, 2H), 3.96 (ddd, 2H), 3.89-3.81 (m, 2H), 3.25-3.19 (m, 2H) , 3.17-3.09(m, 4H), 2.38(dt, 4H), 1.94-1.79(m, 5H), 1.77-1.58(m, 6H), 1.52-1.41(m, 8H), 1.39-1.24(m, 36H), 0.87(t, 6H).
实施例13制备化合物LQ025Example 13 Preparation of Compound LQ025
步骤1：LQ025-1的制备
Step 1: Preparation of LQ025-1
物料配比
Material ratio
操作过程：Operation process:
反应瓶中加入6-溴己酸、3-环戊基-1-丙醇、DCC、DMAP和DCM，室温下搅拌反应12h。TLC(EA:PE＝20:1)显示反应完全。Add 6-bromohexanoic acid, 3-cyclopentyl-1-propanol, DCC, DMAP and DCM to the reaction bottle, and stir for 12 hours at room temperature. TLC (EA:PE=20:1) showed that the reaction was complete.
后处理：Post-processing:
反应液用硅藻土过滤后旋干，柱层析纯化后得2.8g无色油状物。The reaction solution was filtered through diatomaceous earth and spun to dryness. After purification by column chromatography, 2.8 g of colorless oil was obtained.
TLC情况：EA:PE＝20:1，产物Rf＝0.6。TLC situation: EA:PE=20:1, product Rf=0.6.
步骤2：LQ025的制备
Step 2: Preparation of LQ025
物料配比：
Material ratio:
操作过程：Operation process:
反应瓶中加入LQ001-2、LQ025-1、K2CO3、KI、甲基叔丁基醚和乙腈，加热至80℃搅拌反应12h。TLC(DCM:MeOH＝10:1)显示反应完全，产物Rf＝0.4。Add LQ001-2, LQ025-1, K 2 CO 3 , KI, methyl tert-butyl ether and acetonitrile to the reaction bottle, heat to 80°C and stir for 12 hours. TLC (DCM:MeOH=10:1) showed that the reaction was complete and the product Rf=0.4.
后处理：Post-processing:
反应液过滤后旋干，柱层析纯化后得到2g无色油状物，收率为80％。The reaction solution was filtered and spun to dryness. After purification by column chromatography, 2 g of colorless oil was obtained with a yield of 80%.
1HNMR(500MHz，CDCl3)δ：4.86(p，1H)，4.05(t，2H)，3.58(t，2H)，2.64(t，2H)，2.55- 2.46(m，4H)，2.29(dt，4H)，1.75(td，3H)，1.69-1.56(m，8H)，1.55-1.43(m，10H)，1.37-1.28(m，10H)，1.25(s，23H)，1.06(d，3H)，0.87(t，6H)。 1 HNMR (500MHz, CDCl 3 ) δ: 4.86 (p, 1H), 4.05 (t, 2H), 3.58 (t, 2H), 2.64 (t, 2H), 2.55- 2.46(m, 4H), 2.29(dt, 4H), 1.75(td, 3H), 1.69-1.56(m, 8H), 1.55-1.43(m, 10H), 1.37-1.28(m, 10H), 1.25( s, 23H), 1.06 (d, 3H), 0.87 (t, 6H).
LCMS：Rt：3.83min；MS m/z(ESI)：666.7[M+H]+。LCMS: Rt: 3.83min; MS m/z (ESI): 666.7[M+H] + .
实施例14制备化合物LQ026Example 14 Preparation of Compound LQ026
步骤1：LQ026-1的制备
Step 1: Preparation of LQ026-1
物料配比：
Material ratio:
操作过程：Operation process:
反应瓶中加入8-溴辛酸、3-环戊基-1-丙醇、DCC、DMAP和DCM，室温下搅拌反应12h，TLC(EA:PE＝20:1)显示反应完全。Add 8-bromooctanoic acid, 3-cyclopentyl-1-propanol, DCC, DMAP and DCM to the reaction bottle, and stir for 12 hours at room temperature. TLC (EA:PE=20:1) shows that the reaction is complete.
后处理：Post-processing:
反应液用硅藻土过滤后旋干，柱层析纯化后得1.2g无色油状物，收率为65％。The reaction solution was filtered through diatomaceous earth and spun to dryness. After purification by column chromatography, 1.2 g of colorless oil was obtained, with a yield of 65%.
TLC情况：EA:PE＝20:1，产物Rf＝0.6。TLC situation: EA:PE=20:1, product Rf=0.6.
步骤2：LQ026-2的制备
Step 2: Preparation of LQ026-2
物料配比：

Material ratio:

操作过程：Operation process:
反应瓶中加入LQ026-1、乙醇胺和乙腈，室温下搅拌反应12h。TLC(EA:PE＝20:1)显示反应完全。Add LQ026-1, ethanolamine and acetonitrile to the reaction bottle, and stir for 12 hours at room temperature. TLC (EA:PE=20:1) showed that the reaction was complete.
后处理：Post-processing:
反应液旋干后用100mL的乙酸乙酯稀释，用100mL水洗2次，取有机相用无水硫酸钠干燥后旋干，柱层析纯化后得到1g无色油状物，收率为87.8％。The reaction solution was spin-dried, diluted with 100 mL of ethyl acetate, and washed twice with 100 mL of water. The organic phase was dried over anhydrous sodium sulfate and spin-dried. After purification by column chromatography, 1 g of colorless oil was obtained with a yield of 87.8%.
TLC情况：DCM:MeOH＝10:1，产物Rf＝0.2。TLC situation: DCM:MeOH=10:1, product Rf=0.2.
步骤3：LQ026的制备
Step 3: Preparation of LQ026
物料配比：
Material ratio:
操作过程：Operation process:
反应瓶中加入LQ026-2、LQ025-1、K2CO3、KI、甲基叔丁基醚和乙腈，加热至80℃搅拌反应12h。TLC(DCM:MeOH＝10:1)显示反应完全。Add LQ026-2, LQ025-1, K 2 CO 3 , KI, methyl tert-butyl ether and acetonitrile to the reaction bottle, heat to 80°C and stir for 12 hours. TLC (DCM:MeOH=10:1) showed that the reaction was complete.
后处理：Post-processing:
反应液过滤后旋干，柱层析纯化后得到1g无色油状物，收率为76％。The reaction solution was filtered and spun to dryness. After purification by column chromatography, 1 g of colorless oil was obtained with a yield of 76%.
1HNMR(500MHz，CDCl3)δ：4.04(t，4H)，3.58(t，2H)，2.63(t，2H)，2.55-2.47(m，4H)，2.29(td，4H)，1.79-1.70(m，6H)，1.61(ddt，12H)，1.54-1.42(m，8H)，1.37-1.25(m，12H)，1.11-1.01(m，4H)。 1 HNMR (500MHz, CDCl 3 ) δ: 4.04 (t, 4H), 3.58 (t, 2H), 2.63 (t, 2H), 2.55-2.47 (m, 4H), 2.29 (td, 4H), 1.79-1.70 (m, 6H), 1.61 (ddt, 12H), 1.54-1.42 (m, 8H), 1.37-1.25 (m, 12H), 1.11-1.01 (m, 4H).
LCMS：Rt：3.85min；MS m/z(ESI)：538.5[M+H]+。LCMS: Rt: 3.85min; MS m/z (ESI): 538.5[M+H] + .
实施例15制备化合物LQ031Example 15 Preparation of Compound LQ031
步骤1：LQ031-1的制备
Step 1: Preparation of LQ031-1
物料配比
Material ratio
操作过程：Operation process:
反应瓶中加入6-溴己酸、环己基甲醇、DCC、DMAP和DCM，室温下搅拌反应12h，TLC(EA:PE＝20:1)显示反应完全(产物Rf值为0.5)。Add 6-bromohexanoic acid, cyclohexylmethanol, DCC, DMAP and DCM to the reaction bottle, and stir for 12 hours at room temperature. TLC (EA:PE=20:1) shows that the reaction is complete (the product Rf value is 0.5).
后处理：Post-processing:
反应液用硅藻土过滤后旋干，柱层析纯化后得8.6g无色油状物，收率69％。The reaction solution was filtered through diatomaceous earth and spun to dryness. After purification by column chromatography, 8.6 g of colorless oil was obtained, with a yield of 69%.
H NMR(600MHz,CHCl3-d)δ3.88(d,J＝6.6Hz,2H),3.40(t,J＝6.8Hz,2H),2.32(t,J＝7.5Hz,2H),1.88(p,J＝6.9Hz,2H),1.72(d,J＝10.4Hz,4H),1.66(dt,J＝15.5,7.6Hz,4H),1.48(p,J＝7.7Hz,2H),1.29–1.12(m,3H),1.00–0.92(m,2H).H NMR (600MHz, CHCl 3 -d) δ3.88 (d, J = 6.6 Hz, 2H), 3.40 (t, J = 6.8 Hz, 2H), 2.32 (t, J = 7.5 Hz, 2H), 1.88 ( p,J＝6.9Hz,2H),1.72(d,J＝10.4Hz,4H),1.66(dt,J＝15.5,7.6Hz,4H),1.48(p,J＝7.7Hz,2H),1.29– 1.12(m,3H),1.00–0.92(m,2H).
步骤2：LQ031-2的制备
Step 2: Preparation of LQ031-2
物料配比：
Material ratio:
操作过程：Operation process:
反应瓶中加入LQ031-1、乙醇胺和乙腈，加热至30℃搅拌反应12h。TLC(DCM:MeOH＝10:1) 显示反应完全(产物Rf值为0.3)。Add LQ031-1, ethanolamine and acetonitrile to the reaction bottle, heat to 30°C and stir for 12 hours. TLC(DCM:MeOH＝10:1) It shows that the reaction is complete (product Rf value is 0.3).
后处理：Post-processing:
反应液旋干后用500mL的乙酸乙酯稀释，用500mL水洗2次，取有机相用无水硫酸钠干燥后旋干，柱层析纯化后得到5.6g无色油状物。收率69％。The reaction solution was spin-dried, diluted with 500 mL of ethyl acetate, and washed twice with 500 mL of water. The organic phase was dried with anhydrous sodium sulfate and then spin-dried. After purification by column chromatography, 5.6 g of colorless oil was obtained. Yield 69%.
1H NMR(600MHz,CHCl3-d)δ3.87(d,J＝6.6Hz,2H),3.65–3.62(m,2H),2.79–2.74(m,2H),2.65–2.60(m,2H),2.30(t,J＝7.5Hz,2H),1.71(d,J＝10.5Hz,4H),1.68–1.58(m,4H),1.51(p,J＝7.4Hz,2H),1.39–1.32(m,2H),1.28–1.11(m,3H),0.99–0.91(m,2H). 1 H NMR (600MHz, CHCl 3 -d) δ3.87 (d, J＝6.6Hz, 2H), 3.65–3.62 (m, 2H), 2.79–2.74 (m, 2H), 2.65–2.60 (m, 2H) ),2.30(t,J＝7.5Hz,2H),1.71(d,J＝10.5Hz,4H),1.68–1.58(m,4H),1.51(p,J＝7.4Hz,2H),1.39–1.32 (m,2H),1.28–1.11(m,3H),0.99–0.91(m,2H).
步骤3：LQ031的制备
Step 3: Preparation of LQ031
物料配比：
Material ratio:
操作过程：Operation process:
反应瓶中加入LQ031-2、LQ001-1、K2CO3、KI和乙腈，加热至80℃搅拌反应12h。TLC(DCM:MeOH＝10:1)显示反应完全(产物Rf值为0.5)。Add LQ031-2, LQ001-1, K 2 CO 3 , KI and acetonitrile to the reaction bottle, heat to 80°C and stir for 12 hours. TLC (DCM:MeOH=10:1) showed that the reaction was complete (product Rf value was 0.5).
后处理：Post-processing:
反应液过滤后旋干，柱层析纯化后得到1.55g无色油状物。收率64％。The reaction solution was filtered, spun to dryness, and purified by column chromatography to obtain 1.55 g of colorless oil. Yield 64%.
1H NMR(600MHz,CHCl3-d)δ4.86(p,J＝6.8,6.3Hz,1H),3.88(d,J＝6.6Hz,2H),3.56(t,J＝4.9Hz,2H),2.64–2.58(m,2H),2.49(q,J＝6.9Hz,4H),2.29(dt,J＝19.0,7.5Hz,4H),1.72(d,J＝10.2Hz,4H),1.64(tt,J＝14.7,7.2Hz,6H),1.48(dd,J＝21.6,6.1Hz,8H),1.35–1.20(m,34H),1.20–1.14(m,1H),1.00–0.92(m,2H),0.87(t,J＝7.1Hz,6H). 1 H NMR (600MHz, CHCl 3 -d) δ4.86 (p, J＝6.8, 6.3Hz, 1H), 3.88 (d, J＝6.6Hz, 2H), 3.56 (t, J＝4.9Hz, 2H) ,2.64–2.58(m,2H),2.49(q,J＝6.9Hz,4H),2.29(dt,J＝19.0,7.5Hz,4H),1.72(d,J＝10.2Hz,4H),1.64( tt,J＝14.7,7.2Hz,6H),1.48(dd,J＝21.6,6.1Hz,8H),1.35–1.20(m,34H),1.20–1.14(m,1H),1.00–0.92(m, 2H),0.87(t,J=7.1Hz,6H).
实施例16制备化合物LQ032Example 16 Preparation of Compound LQ032
步骤1：LQ032-1的制备
Step 1: Preparation of LQ032-1
物料配比：
Material ratio:
操作过程：Operation process:
反应瓶中加入6-溴己酸、环己基甲醇、DCC、DMAP和DCM，室温下搅拌反应12h，TLC(EA:PE＝20:1)显示反应完全(产物Rf值为0.5)。Add 6-bromohexanoic acid, cyclohexylmethanol, DCC, DMAP and DCM to the reaction bottle, and stir for 12 hours at room temperature. TLC (EA:PE=20:1) shows that the reaction is complete (the product Rf value is 0.5).
后处理：Post-processing:
反应液用硅藻土过滤后旋干，柱层析纯化后得9g无色油状物，收率70％。The reaction solution was filtered through diatomaceous earth and spun to dryness. After purification by column chromatography, 9 g of colorless oil was obtained with a yield of 70%.
步骤2：LQ032的制备
Step 2: Preparation of LQ032
物料配比：
Material ratio:
操作过程：Operation process:
反应瓶中加入LQ031-2、LQ032-1、K2CO3、KI和乙腈，加热至80℃搅拌反应12h。TLC(DCM:MeOH＝10:1)显示反应完全(产物Rf值为0.45)。Add LQ031-2, LQ032-1, K 2 CO 3 , KI and acetonitrile to the reaction bottle, heat to 80°C and stir for 12 hours. TLC (DCM:MeOH=10:1) showed that the reaction was complete (product Rf value was 0.45).
后处理：Post-processing:
反应液过滤后旋干，柱层析纯化后得到1g无色油状物。收率53％。The reaction solution was filtered, spun to dryness, and purified by column chromatography to obtain 1 g of colorless oil. Yield 53%.
1H NMR(600MHz,CHCl3-d)δ3.87(d,J＝6.6Hz,4H),3.70–3.64(m,2H),2.76(s,2H),2.69–2.60(m,4H),2.30(dt,J＝11.1,7.5Hz,4H),1.75–1.52(m,20H),1.37–1.13(m,15H),1.00–0.91(m,4H). 1 H NMR (600MHz, CHCl 3 -d) δ3.87 (d, J＝6.6Hz, 4H), 3.70–3.64 (m, 2H), 2.76 (s, 2H), 2.69–2.60 (m, 4H), 2.30(dt,J＝11.1,7.5Hz,4H),1.75–1.52(m,20H),1.37–1.13(m,15H),1.00–0.91(m,4H).
实施例17制备化合物LQ067 Example 17 Preparation of Compound LQ067
步骤1：LQ067-1的制备
Step 1: Preparation of LQ067-1
物料配比：
Material ratio:
操作过程：Operation process:
反应瓶中加入6-溴己酸、异壬醇、DCC、DMAP和DCM，室温下搅拌反应12h。TLC(EA:PE＝20:1)显示反应完全(产物Rf值为0.6)。Add 6-bromohexanoic acid, isononanol, DCC, DMAP and DCM to the reaction bottle, and stir for 12 hours at room temperature. TLC (EA:PE=20:1) showed that the reaction was complete (product Rf value was 0.6).
后处理：Post-processing:
反应液用硅藻土过滤后旋干，柱层析纯化后得2.4g无色油状物。The reaction solution was filtered through diatomaceous earth and spun to dryness. After purification by column chromatography, 2.4 g of colorless oil was obtained.
步骤2：LQ067的制备
Step 2: Preparation of LQ067
物料配比：
Material ratio:
操作过程：Operation process:
反应瓶中加入LQ001-2、LQ067-1、K2CO3、KI和乙腈，加热至80℃搅拌反应12h。TLC(DCM:MeOH＝10:1)显示反应完全(产Rf值为0.6)。Add LQ001-2, LQ067-1, K 2 CO 3 , KI and acetonitrile to the reaction bottle, heat to 80°C and stir for 12 hours. TLC (DCM:MeOH=10:1) showed that the reaction was complete (yield Rf value was 0.6).
后处理：Post-processing:
反应液过滤后旋干，柱层析纯化后得到1.5g无色油状物。收率71％。 The reaction solution was filtered and spun to dryness. After purification by column chromatography, 1.5 g of colorless oil was obtained. The yield is 71%.
1H NMR(600MHz,CHCl3-d)δ4.84(p,J＝6.3Hz,1H),4.06(t,J＝6.6Hz,2H),3.66(t,J＝5.1Hz,2H),2.76(t,J＝5.0Hz,2H),2.64(q,J＝7.8Hz,4H),2.27(dt,J＝17.6,7.4Hz,4H),1.55(ddtd,J＝53.6,28.1,14.1,13.1,7.0Hz,16H),1.36–1.17(m,35H),0.92(d,J＝6.5Hz,3H),0.88–0.84(m,14H). 1 H NMR (600MHz, CHCl 3 -d) δ4.84 (p, J = 6.3Hz, 1H), 4.06 (t, J = 6.6Hz, 2H), 3.66 (t, J = 5.1Hz, 2H), 2.76 (t,J＝5.0Hz,2H),2.64(q,J＝7.8Hz,4H),2.27(dt,J＝17.6,7.4Hz,4H),1.55(ddtd,J＝53.6,28.1,14.1,13.1 ,7.0Hz,16H),1.36–1.17(m,35H),0.92(d,J=6.5Hz,3H),0.88–0.84(m,14H).
实施例18制备化合物LQ068Example 18 Preparation of Compound LQ068
步骤1：LQ068-1的制备
Step 1: Preparation of LQ068-1
物料配比：
Material ratio:
操作过程：Operation process:
反应瓶中加入8-溴辛酸、异壬醇、DCC、DMAP和DCM，室温下搅拌反应12h，TLC(EA:PE＝20:1)显示反应完全(产物Rf值为0.6)。Add 8-bromooctanoic acid, isononanol, DCC, DMAP and DCM to the reaction bottle, and stir for 12 hours at room temperature. TLC (EA:PE=20:1) shows that the reaction is complete (the product Rf value is 0.6).
后处理：Post-processing:
反应液用硅藻土过滤后旋干，柱层析纯化后得8.8g无色油状物，收率63％。The reaction solution was filtered through diatomaceous earth and spun to dryness. After purification by column chromatography, 8.8 g of colorless oil was obtained, with a yield of 63%.
步骤2：LQ068-2的制备
Step 2: Preparation of LQ068-2
物料配比：
Material ratio:
操作过程：Operation process:
反应瓶中加入LQ068-1、乙醇胺和乙腈，加热至30℃搅拌反应12h。TLC(DCM:MeOH＝10:1)显示反应完全(产物Rf值为0.3)。Add LQ068-1, ethanolamine and acetonitrile to the reaction bottle, heat to 30°C and stir for 12 hours. TLC (DCM:MeOH=10:1) showed that the reaction was complete (product Rf value was 0.3).
后处理：Post-processing:
反应液旋干后用500mL的乙酸乙酯稀释，用500mL水洗2次，取有机相用无水硫酸钠干燥后旋干，柱层析纯化后得到5.8g无色油状物。收率70％。The reaction solution was spin-dried, diluted with 500 mL of ethyl acetate, and washed twice with 500 mL of water. The organic phase was dried over anhydrous sodium sulfate and then spin-dried. After purification by column chromatography, 5.8 g of colorless oil was obtained. The yield is 70%.
步骤3：LQ068的制备
Step 3: Preparation of LQ068
物料配比：
Material ratio:
操作过程：Operation process:
反应瓶中加入LQ068-2、LQ067-1、K2CO3、KI和乙腈，加热至80℃搅拌反应12h。TLC(DCM:MeOH＝10:1)显示反应完全(产物Rf值为0.6)。Add LQ068-2, LQ067-1, K 2 CO 3 , KI and acetonitrile to the reaction bottle, heat to 80°C and stir for 12 hours. TLC (DCM:MeOH=10:1) showed that the reaction was complete (product Rf value was 0.6).
后处理：Post-processing:
反应液过滤后旋干，柱层析纯化后得到1g无色油状物。收率57％。The reaction solution was filtered, spun to dryness, and purified by column chromatography to obtain 1 g of colorless oil. Yield 57%.
1H NMR(600MHz,CHCl3-d)δ4.06(ddd,J＝6.9,4.0,2.5Hz,4H),3.63(t,J＝5.2Hz,2H),2.71(t,J＝5.2Hz,2H),2.59(q,J＝7.6Hz,4H),2.28(dt,J＝9.5,7.5Hz,4H),1.66–1.40(m,15H),1.36–1.19(m,13H),1.06(dd,J＝13.9,6.0Hz,2H),0.93(d,J＝6.5Hz,6H),0.87(s,15H). 1 H NMR (600MHz, CHCl 3 -d) δ4.06 (ddd, J＝6.9, 4.0, 2.5Hz, 4H), 3.63 (t, J＝5.2Hz, 2H), 2.71 (t, J＝5.2Hz, 2H),2.59(q,J＝7.6Hz,4H),2.28(dt,J＝9.5,7.5Hz,4H),1.66–1.40(m,15H),1.36–1.19(m,13H),1.06(dd ,J＝13.9,6.0Hz,2H),0.93(d,J＝6.5Hz,6H),0.87(s,15H).
实施例19制备化合物LQ073Example 19 Preparation of Compound LQ073
步骤1：LQ073-1的制备
Step 1: Preparation of LQ073-1
物料配比：

Material ratio:

操作过程：Operation process:
反应瓶中加入6-溴己酸、8-甲基-1-癸醇、DCC、DMAP和DCM，室温下搅拌反应12h。TLC(EA:PE＝20:1)显示反应完全(产物Rf值为0.6)。Add 6-bromohexanoic acid, 8-methyl-1-decanol, DCC, DMAP and DCM to the reaction bottle, and stir for 12 hours at room temperature. TLC (EA:PE=20:1) showed that the reaction was complete (product Rf value was 0.6).
后处理：Post-processing:
反应液用硅藻土过滤后旋干，柱层析纯化后得2.6g无色油状物。The reaction solution was filtered through diatomaceous earth and spun to dryness. After purification by column chromatography, 2.6 g of colorless oil was obtained.
步骤2：LQ073的制备
Step 2: Preparation of LQ073
物料配比：
Material ratio:
操作过程：Operation process:
反应瓶中加入LQ001-2、LQ073-1、K2CO3、KI和乙腈，加热至80℃搅拌反应12h。TLC(DCM:MeOH＝10:1)显示反应完全(产物Rf值为0.6)。Add LQ001-2, LQ073-1, K 2 CO 3 , KI and acetonitrile to the reaction bottle, heat to 80°C and stir for 12 hours. TLC (DCM:MeOH=10:1) showed that the reaction was complete (product Rf value was 0.6).
后处理：Post-processing:
反应液过滤后旋干，柱层析纯化后得到1.2g无色油状物。收率57％。The reaction solution was filtered and spun to dryness. After purification by column chromatography, 1.2 g of colorless oil was obtained. Yield 57%.
1H NMR(600MHz,CHCl3-d)δ4.84(p,J＝6.2Hz,1H),4.04(t,J＝6.8Hz,2H),3.72(t,J＝5.1Hz,2H),2.82(t,J＝5.1Hz,2H),2.71(q,J＝7.9Hz,3H),2.28(dt,J＝20.3,7.4Hz,4H),1.61(th,J＝14.9,7.1Hz,9H),1.49(q,J＝6.2Hz,4H),1.34–1.21(m,42H),0.88–0.80(m,12H). 1 H NMR (600MHz, CHCl 3 -d) δ4.84 (p, J = 6.2 Hz, 1H), 4.04 (t, J = 6.8 Hz, 2H), 3.72 (t, J = 5.1 Hz, 2H), 2.82 (t,J＝5.1Hz,2H),2.71(q,J＝7.9Hz,3H),2.28(dt,J＝20.3,7.4Hz,4H),1.61(th,J＝14.9,7.1Hz,9H) ,1.49(q,J＝6.2Hz,4H),1.34–1.21(m,42H),0.88–0.80(m,12H).
实施例20制备化合物LQ074 Example 20 Preparation of Compound LQ074
步骤1：LQ074-1的制备
Step 1: Preparation of LQ074-1
物料配比：
Material ratio:
操作过程：Operation process:
反应瓶中加入8-溴辛酸、8-甲基-1-癸醇、DCC、DMAP和DCM，室温下搅拌反应12h，TLC(EA:PE＝20:1)显示反应完全(产物Rf值为0.6)。Add 8-bromooctanoic acid, 8-methyl-1-decanol, DCC, DMAP and DCM to the reaction bottle, and stir for 12 hours at room temperature. TLC (EA:PE=20:1) shows that the reaction is complete (the product Rf value is 0.6 ).
后处理：Post-processing:
反应液用硅藻土过滤后旋干，柱层析纯化后得9.3g无色油状物，收率62％。The reaction solution was filtered through diatomaceous earth and spun to dryness. After purification by column chromatography, 9.3 g of colorless oil was obtained, with a yield of 62%.
步骤2：LQ074-2的制备
Step 2: Preparation of LQ074-2
物料配比：
Material ratio:
操作过程：Operation process:
反应瓶中加入LQ074-1、乙醇胺和乙腈，加热至30℃搅拌反应12h。TLC(DCM:MeOH＝10:1) 显示反应完全(产物Rf值为0.3)。Add LQ074-1, ethanolamine and acetonitrile to the reaction bottle, heat to 30°C and stir for 12 hours. TLC(DCM:MeOH＝10:1) It shows that the reaction is complete (product Rf value is 0.3).
后处理：Post-processing:
反应液旋干后用500mL的乙酸乙酯稀释，用500mL水洗2次，取有机相用无水硫酸钠干燥后旋干，柱层析纯化后得到5.4g无色油状物。收率60％。The reaction solution was spin-dried, diluted with 500 mL of ethyl acetate, and washed twice with 500 mL of water. The organic phase was dried over anhydrous sodium sulfate and then spin-dried. After purification by column chromatography, 5.4 g of colorless oil was obtained. Yield 60%.
步骤3：LQ074的制备
Step 3: Preparation of LQ074
物料配比：
Material ratio:
操作过程：Operation process:
反应瓶中加入LQ074-2、LQ073-1、K2CO3、KI和乙腈，加热至80℃搅拌反应12h。TLC(DCM:MeOH＝10:1)显示反应完全(产物Rf值为0.6)。Add LQ074-2, LQ073-1, K 2 CO 3 , KI and acetonitrile to the reaction bottle, heat to 80°C and stir for 12 hours. TLC (DCM:MeOH=10:1) showed that the reaction was complete (product Rf value was 0.6).
后处理：Post-processing:
反应液过滤后旋干，柱层析纯化后得到1g无色油状物。收率57％。The reaction solution was filtered, spun to dryness, and purified by column chromatography to obtain 1 g of colorless oil. Yield 57%.
1H NMR(600MHz,CHCl3-d)δ4.02(t,J＝6.7Hz,4H),3.61(t,J＝5.3Hz,2H),2.70(t,J＝5.3Hz,2H),2.58(q,J＝7.8Hz,4H),2.26(dt,J＝9.9,7.5Hz,4H),1.64–1.45(m,12H),1.34–1.17(m,31H),1.13–1.01(m,4H),0.84–0.78(m,12H). 1 H NMR (600MHz, CHCl 3 -d) δ4.02 (t, J = 6.7 Hz, 4H), 3.61 (t, J = 5.3 Hz, 2H), 2.70 (t, J = 5.3 Hz, 2H), 2.58 (q,J＝7.8Hz,4H),2.26(dt,J＝9.9,7.5Hz,4H),1.64–1.45(m,12H),1.34–1.17(m,31H),1.13–1.01(m,4H ),0.84–0.78(m,12H).
实施例21制备化合物LQ076Example 21 Preparation of Compound LQ076
步骤1：LQ076-1的制备
Step 1: Preparation of LQ076-1
物料配比：

Material ratio:

操作过程：Operation process:
反应瓶中加入6-溴己酸、3-丁基-1-庚醇、DCC、DMAP和DCM，室温下搅拌反应12h。TLC(EA:PE＝20:1)显示反应完全(产物Rf值为0.6)。Add 6-bromohexanoic acid, 3-butyl-1-heptanol, DCC, DMAP and DCM to the reaction bottle, and stir for 12 hours at room temperature. TLC (EA:PE=20:1) showed that the reaction was complete (product Rf value was 0.6).
后处理：Post-processing:
反应液用硅藻土过滤后旋干，柱层析纯化后得2.8g无色油状物。The reaction solution was filtered through diatomaceous earth and spun to dryness. After purification by column chromatography, 2.8 g of colorless oil was obtained.
步骤2：LQ076的制备
Step 2: Preparation of LQ076
物料配比：
Material ratio:
操作过程：Operation process:
反应瓶中加入LQ001-2、LQ076-1、K2CO3、KI和乙腈，加热至80℃搅拌反应12h。TLC(DCM:MeOH＝10:1)显示反应完全(产物Rf值为0.6)。Add LQ001-2, LQ076-1, K 2 CO 3 , KI and acetonitrile to the reaction bottle, heat to 80°C and stir for 12 hours. TLC (DCM:MeOH=10:1) showed that the reaction was complete (product Rf value was 0.6).
后处理：Post-processing:
反应液过滤后旋干，柱层析纯化后得到1.2g无色油状物。收率57％。The reaction solution was filtered and spun to dryness. After purification by column chromatography, 1.2 g of colorless oil was obtained. Yield 57%.
1H NMR(600MHz,CHCl3-d)δ4.84(p,J＝6.1Hz,1H),4.07(t,J＝7.1Hz,2H),3.66(t,J＝5.2Hz,2H),2.74(t,J＝5.2Hz,2H),2.63(q,J＝7.8Hz,4H),2.27(dt,J＝14.6,7.4Hz,4H),1.66–1.45(m,14H),1.33–1.18(m,43H),0.87(dt,J＝9.5,6.9Hz,12H). 1 H NMR (600MHz, CHCl 3 -d) δ4.84 (p, J = 6.1 Hz, 1H), 4.07 (t, J = 7.1 Hz, 2H), 3.66 (t, J = 5.2 Hz, 2H), 2.74 (t,J＝5.2Hz,2H),2.63(q,J＝7.8Hz,4H),2.27(dt,J＝14.6,7.4Hz,4H),1.66–1.45(m,14H),1.33–1.18( m,43H),0.87(dt,J=9.5,6.9Hz,12H).
实施例22制备化合物LQ077Example 22 Preparation of Compound LQ077
步骤1：LQ077-1的制备
Step 1: Preparation of LQ077-1
物料配比：
Material ratio:
操作过程：Operation process:
反应瓶中加入8-溴辛酸、3-丁基-1-庚醇、DCC、DMAP和DCM，室温下搅拌反应12h，TLC(EA:PE＝20:1)显示反应完全(产物Rf值为0.6)。Add 8-bromooctanoic acid, 3-butyl-1-heptanol, DCC, DMAP and DCM to the reaction bottle, and stir for 12 hours at room temperature. TLC (EA:PE=20:1) shows that the reaction is complete (the product Rf value is 0.6 ).
后处理：Post-processing:
反应液用硅藻土过滤后旋干，柱层析纯化后得9g无色油状物，收率61％。The reaction solution was filtered through diatomaceous earth and spun to dryness. After purification by column chromatography, 9 g of colorless oil was obtained with a yield of 61%.
步骤2：LQ077-2的制备
Step 2: Preparation of LQ077-2
物料配比：
Material ratio:
操作过程：Operation process:
反应瓶中加入LQ068-1、乙醇胺和乙腈，加热至30℃搅拌反应12h。TLC(DCM:MeOH＝10:1)显示反应完全(产物Rf值为0.3)。Add LQ068-1, ethanolamine and acetonitrile to the reaction bottle, heat to 30°C and stir for 12 hours. TLC (DCM:MeOH=10:1) showed that the reaction was complete (product Rf value was 0.3).
后处理： Post-processing:
反应液旋干后用500mL的乙酸乙酯稀释，用500mL水洗2次，取有机相用无水硫酸钠干燥后旋干，柱层析纯化后得到5g无色油状物。收率57％。The reaction solution was spin-dried, diluted with 500 mL of ethyl acetate, and washed twice with 500 mL of water. The organic phase was dried with anhydrous sodium sulfate and then spin-dried. After purification by column chromatography, 5 g of colorless oil was obtained. Yield 57%.
步骤3：LQ077的制备
Step 3: Preparation of LQ077
物料配比：
Material ratio:
操作过程：Operation process:
反应瓶中加入LQ077-2、LQ076-1、K2CO3、KI和乙腈，加热至80℃搅拌反应12h。TLC(DCM:MeOH＝10:1)显示反应完全(产物Rf值为0.6)。Add LQ077-2, LQ076-1, K 2 CO 3 , KI and acetonitrile to the reaction bottle, heat to 80°C and stir for 12 hours. TLC (DCM:MeOH=10:1) showed that the reaction was complete (product Rf value was 0.6).
后处理：Post-processing:
反应液过滤后旋干，柱层析纯化后得到1.1g无色油状物。收率57％。The reaction solution was filtered, spun to dryness, and purified by column chromatography to obtain 1.1 g of colorless oil. Yield 57%.
1H NMR(600MHz,CHCl3-d)δ4.07(t,J＝7.1Hz,4H),3.56(dt,J＝10.7,5.3Hz,2H),2.61(dt,J＝13.5,5.3Hz,2H),2.50(p,J＝7.4Hz,4H),2.28(q,J＝7.4Hz,4H),1.66–1.54(m,8H),1.52–1.43(m,4H),1.38(q,J＝6.2Hz,2H),1.35–1.19(m,32H),0.88(t,J＝6.9Hz,12H). 1 H NMR (600MHz, CHCl 3 -d) δ4.07 (t, J = 7.1 Hz, 4H), 3.56 (dt, J = 10.7, 5.3 Hz, 2H), 2.61 (dt, J = 13.5, 5.3 Hz, 2H),2.50(p,J＝7.4Hz,4H),2.28(q,J＝7.4Hz,4H),1.66–1.54(m,8H),1.52–1.43(m,4H),1.38(q,J ＝6.2Hz,2H),1.35–1.19(m,32H),0.88(t,J＝6.9Hz,12H).
实施例23脂质纳米颗粒(LNP)的制备和检测Example 23 Preparation and detection of lipid nanoparticles (LNP)
将系列可电离脂质化合物、二硬脂酰基磷脂酰胆碱(DSPC，日本精化株式会社，货号：S01005)、胆固醇(日本精化株式会社，货号：O01001)和二肉豆蔻酰甘油-聚乙二醇2000(DMG-PEG2000，国邦药业，货号：O02005)分别溶于乙醇溶液，继而依据一定的摩尔比例混合制得混合脂质的乙醇溶液(脂质总浓度为12.5mM)。将萤火虫荧光素酶(Fluc)mRNA或SARS-CoV-2刺突蛋白(Spike)mRNA(SARS-CoV-2刺突蛋白mRNA参见Tan,S.等人，bioRxiv 2022.05.10.491301.)在50mM柠檬酸盐缓冲液(pH 4.0)中稀释得到mRNA溶液。通过使用微流控装置以流速12mL/min、体积比1：3混合脂质的乙醇溶液和mRNA溶液，以可电离脂质与mRNA的氮磷比为6：1制备脂质纳米颗粒。经0.01M的磷酸缓冲液(PBS)透析12-24h除去乙醇。最后，脂质纳米颗粒溶液通过0.22μm无菌过滤器过滤，并经超滤浓缩(Amicon-Ultra，MWCO 10KDa)得到使用可电离脂质/DSPC/胆固醇 /DMG-PEG2000包封Fluc mRNA或Spike mRNA的LNP制剂。SM102可电离脂质(浙江神洲药业有限公司，批号：W211-YB211202)做阳性对照，其结构如下：A series of ionizable lipid compounds, distearoylphosphatidylcholine (DSPC, Nippon Seika Co., Ltd., product number: S01005), cholesterol (Nihon Seika Co., Ltd., product number: O01001) and dimyristoylglycerol-poly Ethylene glycol 2000 (DMG-PEG2000, Guobang Pharmaceutical, product number: O02005) was dissolved in ethanol solution, and then mixed according to a certain molar ratio to prepare an ethanol solution of mixed lipids (total lipid concentration is 12.5mM). Firefly luciferase (Fluc) mRNA or SARS-CoV-2 spike protein (Spike) mRNA (for SARS-CoV-2 spike protein mRNA, see Tan, S. et al., bioRxiv 2022.05.10.491301.) were dissolved in 50mM citric acid Dilute in salt buffer (pH 4.0) to obtain an mRNA solution. Lipid nanoparticles were prepared by using a microfluidic device to mix the ethanol solution of lipid and the mRNA solution at a flow rate of 12 mL/min and a volume ratio of 1:3, with a nitrogen-phosphorus ratio of ionizable lipid to mRNA of 6:1. The ethanol was removed by dialysis against 0.01 M phosphate buffered saline (PBS) for 12-24 h. Finally, the lipid nanoparticle solution was filtered through a 0.22 μm sterile filter and concentrated by ultrafiltration (Amicon-Ultra, MWCO 10 KDa) to obtain ionizable lipid/DSPC/cholesterol. /DMG-PEG2000 LNP preparation encapsulating Fluc mRNA or Spike mRNA. SM102 ionizable lipid (Zhejiang Shenzhou Pharmaceutical Co., Ltd., batch number: W211-YB211202) was used as a positive control. Its structure is as follows:
无特殊说明，可电离脂质/DSPC/胆固醇/DMG-PEG2000的摩尔比为50：10：38.5：1.5。使用Malvern Zetasizer Ultra(动态光散射法)测定各LNP的粒径大小及多分散指数(PDI)；使用Quant-it Ribogreen RNA定量测定试剂盒(ThermoFisher Scientific，货号：R11490)测定LNP的包封效率；6-(p-Toluidino)-2-naphthalene sulfonic acid sodium salt(TNS，南京熙泽医药科技有限公司，货号：XZ0743)染料结合试验测量LNP的pKa，核酸凝胶电泳考察mRNA完整性，测试结果见表1及图1(各Fluc-mRNA LNP凝胶电泳图)。 Without special instructions, the molar ratio of ionizable lipid/DSPC/cholesterol/DMG-PEG2000 is 50:10:38.5:1.5. Malvern Zetasizer Ultra (dynamic light scattering method) was used to measure the particle size and polydispersity index (PDI) of each LNP; Quant-it Ribogreen RNA quantitative assay kit (ThermoFisher Scientific, Cat. No.: R11490) was used to measure the encapsulation efficiency of LNP; 6-(p-Toluidino)-2-naphthalene sulfonic acid sodium salt (TNS, Nanjing Xize Pharmaceutical Technology Co., Ltd., product number: XZ0743) dye binding test measured the pKa of LNP, and nucleic acid gel electrophoresis was used to examine the integrity of the mRNA. The test results are shown in Table 1 and Figure 1 (each Fluc-mRNA LNP gel electrophoresis pattern).
表1 各LNP的粒径、PDI、包封率及pKa(n＝3)。
Table 1 Particle size, PDI, encapsulation efficiency and pKa of each LNP (n=3).
在本领域中，PDI小于0.3，则说明该LNP制剂中纳米颗粒大小相对均匀；包封率用来说明LNP是否能有效包载mRNA，包封率高于70％，则说明该LNP可有效包载mRNA；琼脂糖凝胶电泳图中条带单一且明亮可说明mRNA结构完整。其中，PDI越趋于0越好，包封率越趋于100％越好。由表1可见，实施例23制备的各LNP粒径在60～150nm之间，PDI均小于0.2，各可电离脂质对应的LNP pKa均大于5.5。由图1可见，SM102、LQ001、LQ007、LQ013、LQ016、LQ025、LQ017对应的各mRNA-LNP电泳后mRNA条带清晰明亮，表明其均可有效包载mRNA并维持mRNA结构完整；LQ109对应的LNP的孔道未见明亮条带，表明该组mRNA完整性欠佳。 In this field, if the PDI is less than 0.3, it means that the size of the nanoparticles in the LNP preparation is relatively uniform; the encapsulation rate is used to indicate whether the LNP can effectively encapsulate mRNA. The encapsulation rate is higher than 70%, which means that the LNP can effectively encapsulate mRNA. Loading mRNA; single and bright bands in the agarose gel electrophoresis diagram indicate that the mRNA structure is complete. Among them, the closer the PDI is to 0, the better, and the closer the encapsulation rate is to 100%, the better. It can be seen from Table 1 that the particle size of each LNP prepared in Example 23 is between 60 and 150 nm, the PDI is less than 0.2, and the LNP pKa corresponding to each ionizable lipid is greater than 5.5. As can be seen from Figure 1, the mRNA bands of each mRNA-LNP corresponding to SM102, LQ001, LQ007, LQ013, LQ016, LQ025, and LQ017 are clear and bright after electrophoresis, indicating that they can effectively encapsulate mRNA and maintain the integrity of the mRNA structure; the LNP corresponding to LQ109 There are no bright bands in the channels, indicating that the integrity of the mRNA in this group is poor.
实施例24脂质纳米颗粒(LNP)的制备和检测Example 24 Preparation and detection of lipid nanoparticles (LNP)
采用与实施例23相同的制备方式制得8种LNP制剂(包载Fluc mRNA)，并测定它们的粒径、PDI及包封率。其中，以可电离脂质与mRNA的氮磷比为6：1制备脂质纳米颗粒，可电离脂质/DSPC/胆固醇/DMG-PEG2000的摩尔比为50：10：38.5：1.5，以四个组分的总摩尔数为100％计，各组制剂摩尔百分含量一致。Eight kinds of LNP preparations (encapsulated Fluc mRNA) were prepared using the same preparation method as Example 23, and their particle size, PDI and encapsulation efficiency were measured. Among them, lipid nanoparticles were prepared with a nitrogen-phosphorus ratio of ionizable lipid to mRNA of 6:1, and a molar ratio of ionizable lipid/DSPC/cholesterol/DMG-PEG2000 of 50:10:38.5:1.5, with four The total number of moles of the components is 100%, and the molar percentages of each group of preparations are consistent.
表2 各LNP的粒径、PDI及包封率
Table 2 Particle size, PDI and encapsulation efficiency of each LNP
由表2可见，由上述不同可电离脂质制得的各组LNP制剂粒径介于60～250nm之间；PDI均小于0.25，具体介于0.038至0.234；包封率均高于70％。It can be seen from Table 2 that the particle size of each group of LNP preparations prepared from the above different ionizable lipids ranges from 60 to 250 nm; the PDI is less than 0.25, specifically between 0.038 and 0.234; the encapsulation rate is higher than 70%.
实施例25 LNP制剂的体外细胞实验Example 25 In vitro cell experiments of LNP preparations
以每孔1万个293FT细胞铺种至96孔板，过夜培养至细胞贴壁。将实施例23中制备的Fluc-mRNA LNP制剂(按100ng mRNA/孔计)分别加入96孔板的细胞培养液中(替换为无抗生素、含10％胎牛血清(FBS)的DMEM培养基)(厂家：Gibco，货号：C11995500BT)，继续培养18h后弃去细胞上清，100μL细胞裂解液裂解细胞，加D-荧光素钾盐(PerkinElmer，货号：122799，终浓度1mM)和ATP(厂家：ApexBio，货号：C6931，终浓度2mM)，酶标仪检测化学发光强度，测试结果见图2(293FT细胞与各Fluc-mRNA LNP共培养18-24h后的化学发光强度。通过ANOVA进行统计学分析。***p＜0.001，与PBS组比较)。通过体外细胞实验来验证本申请LNP制剂的体外细胞递送及表达，测试结果见图2。10,000 293FT cells per well were seeded into a 96-well plate and cultured overnight until the cells adhered. The Fluc-mRNA LNP preparations prepared in Example 23 (based on 100ng mRNA/well) were added to the cell culture medium of the 96-well plate (replaced with DMEM culture medium without antibiotics and containing 10% fetal bovine serum (FBS)). (Manufacturer: Gibco, Catalog No.: C11995500BT). After continuing to culture for 18 hours, discard the cell supernatant, lyse the cells with 100 μL of cell lysis solution, and add D-fluorescein potassium salt (PerkinElmer, Catalog No.: 122799, final concentration 1mM) and ATP (Manufacturer: ApexBio, product number: C6931, final concentration 2mM), the chemiluminescence intensity was detected by a microplate reader. The test results are shown in Figure 2 (chemiluminescence intensity of 293FT cells after co-culture with each Fluc-mRNA LNP for 18-24 hours. Statistical analysis was performed by ANOVA .***p<0.001, compared with PBS group). The in vitro cell delivery and expression of the LNP preparation of this application were verified through in vitro cell experiments. The test results are shown in Figure 2.
在图2中，PBS为阴性对照，此组对应的化学发光强度读数可视为背景读值。化学发光强度读数越高，说明表达的越高。由图2可见，LQ001、LQ007、LQ013、LQ016及LQ025对应的Fluc-mRNA  LNP均可有效递送Fluc mRNA进入细胞并表达。In Figure 2, PBS is a negative control, and the chemiluminescence intensity readings corresponding to this group can be regarded as background readings. The higher the chemiluminescence intensity reading, the higher the expression. As can be seen from Figure 2, the Fluc-mRNA corresponding to LQ001, LQ007, LQ013, LQ016 and LQ025 Both LNPs can effectively deliver Fluc mRNA into cells and express it.
以每孔40万个293FT细胞铺种至12孔板，过夜培养至细胞贴壁。将实施例23中制备的Spike-mRNA LNP(按1μg/mL/孔计)分别加入12孔板的细胞培养液中(替换为无抗生素、含10％胎牛血清(FBS)的DMEM培养基)(厂家：Gibco，货号：C11995500BT)，继续培养24h。裂解细胞并提取总蛋白，Western blot法测定Spike蛋白的细胞水平表达情况，测试结果见图3(293FT细胞与各Spike-mRNA LNP共培养24h后的Spike蛋白表达情况，PBS组作为阴性对照)。400,000 293FT cells per well were seeded into a 12-well plate and cultured overnight until the cells adhered. The Spike-mRNA LNPs prepared in Example 23 (based on 1 μg/mL/well) were added to the cell culture medium of the 12-well plate (replaced with DMEM culture medium without antibiotics and containing 10% fetal bovine serum (FBS)). (Manufacturer: Gibco, Product No.: C11995500BT), continue culturing for 24 hours. The cells were lysed and total protein was extracted. The cellular level expression of Spike protein was determined by Western blot method. The test results are shown in Figure 3 (Spike protein expression after 293FT cells were co-cultured with each Spike-mRNA LNP for 24 hours, and the PBS group was used as a negative control).
由图3可见，LQ001、LQ007、LQ013、LQ025对应的Spike-mRNA LNP制剂均可有效递送Spike mRNA进入细胞并表达。As can be seen from Figure 3, the Spike-mRNA LNP preparations corresponding to LQ001, LQ007, LQ013, and LQ025 can effectively deliver Spike mRNA into cells and express it.
实施例26 LNP制剂的动物体内研究Example 26 In vivo animal studies of LNP formulations
按5μg/只的剂量将实施例23制备的Fluc-mRNA-LNP通过尾静脉注射到6-8周龄雌性Balb/C小鼠(维通利华)中，并在给药后特定的时间节点(本实施例中为6、24、48、72h)腹腔注射D-荧光素钾盐，然后经IVIS Spectrum小动物活体成像仪检测LNP所携带的mRNA在小鼠体内表达的蛋白质的辐射强度(对应于生物发光表达强度)，统计小鼠活体肝脏部位总发光强度。测试结果见图4(静脉注射给药各Fluc-mRNA LNP不同时间后的体内总生物发光量(n＝3)，PBS组做阴性对照(n＝2))。规定时间点，腹腔内注射荧光素10min后，通过生物发光成像测量荧光素酶活性，Living Image软件(PerkinElmer)统计肝脏区域的总发光强度。进一步，通过GraphPad软件计算曲线下面积(AUC，单位：p/s*小时)，即药后第4小时或第6小时(采用本申请的实验方法，药后3-6小时的峰值相同)至药后第72小时总发光强度测量点连线的曲线下面积，AUC越大，说明表达越好。The Fluc-mRNA-LNP prepared in Example 23 was injected into 6-8 week old female Balb/C mice (Vital Lever) through the tail vein at a dose of 5 μg/mouse, and at a specific time point after administration (6, 24, 48, 72 hours in this example) D-luciferin potassium salt was injected intraperitoneally, and then the radiation intensity of the protein expressed by the mRNA carried by the LNP in the mouse was detected by the IVIS Spectrum small animal live imager (corresponding to (bioluminescence expression intensity), and the total luminescence intensity of the mouse liver in vivo was calculated. The test results are shown in Figure 4 (the total bioluminescence amount in the body after intravenous injection of each Fluc-mRNA LNP at different times (n=3), the PBS group was used as a negative control (n=2)). At specified time points, 10 minutes after intraperitoneal injection of luciferin, luciferase activity was measured through bioluminescence imaging, and Living Image software (PerkinElmer) counted the total luminescence intensity in the liver area. Further, calculate the area under the curve (AUC, unit: p/s*hour) through GraphPad software, that is, the 4th or 6th hour after the drug (using the experimental method of this application, the peak value at 3-6 hours after the drug is the same) to The area under the curve connecting the total luminescence intensity measurement points at 72 hours after taking the drug. The larger the AUC, the better the expression.
如图4所示，LQ001、LQ007、LQ013、LQ016和LQ025对应的Fluc-mRNA LNP制剂在小鼠体内均有较强表达，一直到72h仍能够观察到比本底水平(PBS组)高10倍以上的发光强度。As shown in Figure 4, the Fluc-mRNA LNP preparations corresponding to LQ001, LQ007, LQ013, LQ016 and LQ025 all have strong expression in mice, and can still be observed to be 10 times higher than the background level (PBS group) until 72h. luminous intensity above.
按2μg/只的剂量将实施例23制备的Spike-mRNA-LNP制剂通过肌肉注射到6-8周龄雌性Balb/C小鼠(维通利华)中，在第一次注射后21天重复注射相应的mRNA-LNP制剂。并在2次给药后特定的时间节点(本次实施例中数据为第二次给药后一周)采集小鼠全血。4℃、2000g离心10min从全血中分离出血清，血清在56℃水浴灭活30min，-80℃下保存以用于分析。根据制造商的说明，使用SARS-CoV-2(2019-nCoV))Spike S1+S2ECD-His Recombinant Protein(义翘神州，货号：40589-V08B1)包被抗原，用SARS-CoV-2(2019-nCoV)Spike Neutralizing Antibody Mouse Mab(义翘神州，货号：40591-MM43)做对照，2％BSA封闭，Peroxidase AffiniPure Goat Anti-Mouse IgG(H+L)(Jackson ImmunoResearch，货号：115-035-003)二抗孵育，按照说明书，使用TMB(Invitrogen，货号：00-4201-56)显色进行酶联免疫吸附测定(ELISA)分析，测得Spike抗体滴度，测试结果见图5(末次给药后1周小鼠血清中的总抗体滴度(n＝8)。通过ANOVA进行统计学分析。**p＜0.01，***p＜0.001，与PBS组比较)。The Spike-mRNA-LNP preparation prepared in Example 23 was injected intramuscularly into 6-8 week old female Balb/C mice (Vital Lever) at a dose of 2 μg/mouse, and repeated 21 days after the first injection. Inject the corresponding mRNA-LNP preparation. And at a specific time point after the second administration (the data in this example is one week after the second administration), the whole blood of the mice was collected. Serum was separated from whole blood by centrifugation at 2000 g for 10 min at 4°C. The serum was inactivated in a water bath at 56°C for 30 min and stored at -80°C for analysis. According to the manufacturer's instructions, use SARS-CoV-2 (2019-nCoV)) Spike S1+S2ECD-His Recombinant Protein (Yiqiao Shenzhou, Cat. No.: 40589-V08B1) to coat the antigen, and use SARS-CoV-2 (2019- nCoV) Spike Neutralizing Antibody Mouse Mab (Yiqiao Shenzhou, Cat. No.: 40591-MM43) as control, 2% BSA blocked, Peroxidase AffiniPure Goat Anti-Mouse IgG (H+L) (Jackson ImmunoResearch, Cat. No.: 115-035-003) Incubate with the secondary antibody, and use TMB (Invitrogen, Cat. No.: 00-4201-56) to develop color and perform enzyme-linked immunosorbent assay (ELISA) analysis according to the instructions. The Spike antibody titer is measured. The test results are shown in Figure 5 (after the last dose). Total antibody titers in serum of mice at 1 week (n=8). Statistical analysis was performed by ANOVA. **p<0.01, ***p<0.001, compared with PBS group).
与PBS组动物相比，SM102、LQ001、LQ013、LQ025、LQ007、LQ010、LQ016组动物Spike蛋白总抗体滴度有明显的增加，而LQ002、LQ014、LQ109组未见增加。Compared with the animals in the PBS group, the total antibody titer of Spike protein in the animals in the SM102, LQ001, LQ013, LQ025, LQ007, LQ010, and LQ016 groups increased significantly, while there was no increase in the LQ002, LQ014, and LQ109 groups.
实施例27 LNP制剂的动物体内研究 Example 27 In vivo animal studies of LNP formulations
参考实施例26的小鼠体内试验方法，按5μg/只的剂量将该实施例24制备的所有LNP制剂通过尾静脉注射到6至8周龄的雌性Balb/C小鼠体内，并在给药后特定的时间节点(本实施例中为4、24、48h)腹腔注射D-荧光素钾盐，接下来采用与实施例26相同的方法统计肝脏区域的总发光强度，测试结果见图6。实验结果表明：其中5组(LQ031、LQ067、LQ073、LQ076、LQ077)LNP制剂在小鼠体内有较强表达，表明其体内递送能力较好。 Referring to the mouse in vivo test method of Example 26, all the LNP preparations prepared in Example 24 were injected into 6 to 8-week-old female Balb/C mice through the tail vein at a dose of 5 μg/mouse, and after administration D-fluorescein potassium salt was injected intraperitoneally at specific time points (4, 24, and 48 hours in this example), and then the total luminescence intensity of the liver region was calculated using the same method as in Example 26. The test results are shown in Figure 6. The experimental results showed that 5 groups of LNP preparations (LQ031, LQ067, LQ073, LQ076, and LQ077) had strong expression in mice, indicating their better in vivo delivery capabilities.

Claims (16)

  1. 一种如式I所示的含氮链状化合物或其药学上可接受的盐，其特征在于，
    A nitrogen-containing chain compound represented by formula I or a pharmaceutically acceptable salt thereof, characterized in that,
    X为C1-C6的亚烷基或C3-C6的亚环烷基；X is a C 1 -C 6 alkylene group or a C 3 -C 6 cycloalkylene group;
    Y为未取代的或被1个、2个或3个Y1-1取代C1-C10的亚烷基；Y is an unsubstituted or C 1 -C 10 alkylene group substituted by 1, 2 or 3 Y 1-1 ;
    各个Y1-1独立地为羟基、卤素或C1-C6的烷基；Each Y 1-1 is independently hydroxyl, halogen or C 1 -C 6 alkyl;
    Z为未取代的或被1个、2个或3个Z1-1取代C1-C10的亚烷基；Z is an unsubstituted or C 1 -C 10 alkylene group substituted by 1, 2 or 3 Z 1-1 ;
    各个Z1-1独立地为羟基、卤素或C1-C6的烷基；Each Z 1-1 is independently hydroxyl, halogen or C 1 -C 6 alkyl;
    A1和A2独立地为 A 1 and A 2 are independently
    R1为未取代的或被1个、2个或3个R1-1取代的C1-C20的烷基或未取代的或被1个、2个或3个R1-2取代的C2-C20的烯基；R 1 is a C 1 -C 20 alkyl group that is unsubstituted or substituted by 1, 2 or 3 R 1-1 or unsubstituted or substituted by 1, 2 or 3 R 1-2 C 2 -C 20 alkenyl;
    各个R1-1独立地为羟基、C1-C10的烷基、未取代的或被1个、2个或3个R1-1-4取代的C3-C15的环烷基、 Each R 1-1 is independently a hydroxyl group, a C 1 -C 10 alkyl group, a C 3 -C 15 cycloalkyl group that is unsubstituted or substituted by 1, 2 or 3 R 1-1-4 ,
    R1-1-1为C1-C6的烷基；R1-1-2为C1-C6的烷基；R1-1-3为C1-C6的烷基；各个R1-1-4独立地为C1-C6的烷基；R 1-1-1 is a C 1 -C 6 alkyl group; R 1-1-2 is a C 1 -C 6 alkyl group; R 1-1-3 is a C 1 -C 6 alkyl group; each R 1-1-4 is independently a C 1 -C 6 alkyl group;
    各个R1-2独立地为羟基或C1-C10的烷基；Each R 1-2 is independently a hydroxyl group or a C 1 -C 10 alkyl group;
    R2为未取代的或被1个、2个或3个R2-1取代的C1-C20的烷基或未取代的或被1个、2个或3个R2-2取代的C2-C20的烯基；R 2 is a C 1 -C 20 alkyl group that is unsubstituted or substituted by 1, 2 or 3 R 2-1 or unsubstituted or substituted by 1, 2 or 3 R 2-2 C 2 -C 20 alkenyl;
    各个R2-1独立地为羟基、C1-C10的烷基、未取代的或被1个、2个或3个R2-1-4取代的C3-C15的环烷基、 Each R 2-1 is independently a hydroxyl group, a C 1 -C 10 alkyl group, a C 3 -C 15 cycloalkyl group that is unsubstituted or substituted by 1, 2 or 3 R 2-1-4 ,
    R2-1-1为C1-C6的烷基；R2-1-2为C1-C6的烷基；R2-1-3为C1-C6的烷基；各个R2-1-4独立地为C1-C6的烷基；R 2-1-1 is a C 1 -C 6 alkyl group; R 2-1-2 is a C 1 -C 6 alkyl group; R 2-1-3 is a C 1 -C 6 alkyl group; each R 2-1-4 is independently a C 1 -C 6 alkyl group;
    各个R2-2独立地为羟基或C1-C10的烷基：Each R 2-2 is independently hydroxyl or C 1 -C 10 alkyl:
    当X为C1-C6的亚烷基时，R1为被1个、2个或3个羟基取代的C1-C20的烷基、未取代的或被1个、2个或3个R1-1-4取代的环戊烷、未取代的或被1个、2个或3个R1-1-4取代的环12烷、 When X is a C 1 -C 6 alkylene group, R 1 is a C 1 -C 20 alkyl group substituted by 1, 2 or 3 hydroxyl groups, unsubstituted, or Cyclopentane substituted with R 1-1-4 , cyclopentane unsubstituted or substituted by 1, 2 or 3 R 1-1-4 ,
  2. 如权利要求1所述的如式I所示的含氮链状化合物或其药学上可接受的盐，其特征在于，所述如式I所示的含氮链状化合物满足如下条件中的一种或多种：The nitrogen-containing chain compound represented by formula I or a pharmaceutically acceptable salt thereof according to claim 1, characterized in that the nitrogen-containing chain compound represented by formula I satisfies one of the following conditions: One or more species:
    (1)X中，所述C1-C6的亚烷基为C1-C4的亚烷基，优选 例如 (1) In X, the C 1 -C 6 alkylene group is a C 1 -C 4 alkylene group, preferably For example
    (2)X中，所述C3-C6的亚环烷基为 例如 (2) In X, the C 3 -C 6 cycloalkylene group is For example
    (3)Y中，所述C1-C10的亚烷基为C4-C6的亚烷基，优选 例如 (3) In Y, the C 1 -C 10 alkylene group is a C 4 -C 6 alkylene group, preferably For example
    (4)各个Y1-1中，所述卤素为氟、氯、溴或碘；(4) In each Y 1-1 , the halogen is fluorine, chlorine, bromine or iodine;
    (5)各个Y1-1中，所述C1-C6的亚烷基为 (5) In each Y 1-1 , the C 1 -C 6 alkylene group is
    (6)Z中，所述C1-C10的亚烷基为C5-C8的亚烷基，优选 例如 (6) In Z, the C 1 -C 10 alkylene group is a C 5 -C 8 alkylene group, preferably For example
    (7)各个Z1-1中，所述卤素为氟、氯、溴或碘；(7) In each Z 1-1 , the halogen is fluorine, chlorine, bromine or iodine;
    (8)各个Z1-1中，所述C1-C6的亚烷基为 (8) In each Z 1-1 , the C 1 -C 6 alkylene group is
    (9)R1中，所述C1-C20的烷基为C1-C15的烷基，优选为直链烷基，例如乙基、丙基、己基、辛基、壬基或十二烷基；(9) In R 1 , the C 1 -C 20 alkyl group is a C 1 -C 15 alkyl group, preferably a straight-chain alkyl group, such as ethyl, propyl, hexyl, octyl, nonyl or decyl. dialkyl;
    (10)R1中，所述C2-C20的烯基为C6-C18的烯基，优选为直链烯烃基；所述C2-C20的烯基可含有1到4个双键；(10) In R 1 , the C 2 -C 20 alkenyl group is a C 6 -C 18 alkenyl group, preferably a linear alkenyl group; the C 2 -C 20 alkenyl group may contain 1 to 4 double bond;
    (11)各个R1-1中，所述C1-C10的烷基为C1-C4的烷基，优选甲基、乙基、正丙基、异丙基、正丁基、叔丁基、异丁基或仲丁基，例如甲基、乙基、异丙基、叔丁基或正丁基；(11) In each R 1-1 , the C 1 -C 10 alkyl group is a C 1 -C 4 alkyl group, preferably methyl, ethyl, n-propyl, isopropyl, n-butyl, tert. Butyl, isobutyl or sec-butyl, such as methyl, ethyl, isopropyl, tert-butyl or n-butyl;
    (12)各个R1-1中，所述C3-C15的环烷基为C3-C12的环烷基，优选C4-C8的环烷基或环12烷基，例如环戊基、环己基或环12烷基；(12) In each R 1-1 , the C 3 -C 15 cycloalkyl group is a C 3 -C 12 cycloalkyl group, preferably a C 4 -C 8 cycloalkyl group or a cyclo12 alkyl group, such as cycloalkyl group Pentyl, cyclohexyl or cyclo12alkyl;
    (13)R1-1-1中，所述C1-C6的烷基为C1-C4的烷基，优选甲基、乙基、正丙基、异丙基、正丁基、 叔丁基、异丁基或仲丁基；(13) In R 1-1-1 , the C 1 -C 6 alkyl group is a C 1 -C 4 alkyl group, preferably methyl, ethyl, n-propyl, isopropyl, n-butyl, tert-butyl, isobutyl or sec-butyl;
    (14)R1-1-2中，所述C1-C6的烷基为C1-C4的烷基，优选甲基、乙基、正丙基、异丙基、正丁基、叔丁基、异丁基或仲丁基；(14) In R 1-1-2 , the C 1 -C 6 alkyl group is a C 1 -C 4 alkyl group, preferably methyl, ethyl, n-propyl, isopropyl, n-butyl, tert-butyl, isobutyl or sec-butyl;
    (15)R1-1-3中，所述C1-C6的烷基为C1-C4的烷基，优选甲基、乙基、正丙基、异丙基、正丁基、叔丁基、异丁基或仲丁基；(15) In R 1-1-3 , the C 1 -C 6 alkyl group is a C 1 -C 4 alkyl group, preferably methyl, ethyl, n-propyl, isopropyl, n-butyl, tert-butyl, isobutyl or sec-butyl;
    (16)各个R1-1-4中，所述C1-C6的烷基为C1-C4的烷基，优选甲基、乙基、正丙基、异丙基、正丁基、叔丁基、异丁基或仲丁基；(16) In each R 1-1-4 , the C 1 -C 6 alkyl group is a C 1 -C 4 alkyl group, preferably methyl, ethyl, n-propyl, isopropyl, n-butyl , tert-butyl, isobutyl or sec-butyl;
    (17)各个R1-2中，所述C1-C10的烷基为C1-C6的烷基，优选甲基、乙基、正丙基、异丙基、正丁基、叔丁基、异丁基或仲丁基；(17) In each R 1-2 , the C 1 -C 10 alkyl group is a C 1 -C 6 alkyl group, preferably methyl, ethyl, n-propyl, isopropyl, n-butyl, tert. Butyl, isobutyl or sec-butyl;
    (18)R2中，所述C1-C20的烷基为C1-C15的烷基，优选为直链烷基，例如乙基、丙基、己基、辛基、壬基或十二烷基；(18) In R 2 , the C 1 -C 20 alkyl group is a C 1 -C 15 alkyl group, preferably a straight chain alkyl group, such as ethyl, propyl, hexyl, octyl, nonyl or decyl. dialkyl;
    (19)R2中，所述C2-C20的烯基为C6-C18的烯基，优选为直链烯基；所述C2-C20的烯基可含有1到4个双键；(19) In R 2 , the C 2 -C 20 alkenyl group is a C 6 -C 18 alkenyl group, preferably a linear alkenyl group; the C 2 -C 20 alkenyl group may contain 1 to 4 double bond;
    (20)各个R2-1中，所述C1-C10的烷基为C1-C8的烷基，优选甲基、乙基、正丙基、异丙基、正丁基、叔丁基、异丁基、仲丁基、正戊基、异戊基、正己基、异己基、正庚基、异庚基、正辛基或异辛基，例如甲基、乙基、异丙基、叔丁基、正丁基或正辛基；(20) In each R 2-1 , the C 1 -C 10 alkyl group is a C 1 -C 8 alkyl group, preferably methyl, ethyl, n-propyl, isopropyl, n-butyl, tert. Butyl, isobutyl, sec-butyl, n-pentyl, isopentyl, n-hexyl, isohexyl, n-heptyl, isoheptyl, n-octyl or isooctyl, e.g. methyl, ethyl, isopropyl base, tert-butyl, n-butyl or n-octyl;
    (21)各个R2-1中，所述C3-C15的环烷基为C3-C12的环烷基，优选C4-C8的环烷基或环12烷基，例如环戊基、环己基或环12烷基；(21) In each R 2-1 , the C 3 -C 15 cycloalkyl group is a C 3 -C 12 cycloalkyl group, preferably a C 4 -C 8 cycloalkyl group or a cyclo12 alkyl group, such as cycloalkyl group Pentyl, cyclohexyl or cyclo12alkyl;
    (22)R2-1-1中，所述C1-C6的烷基为C1-C4的烷基，优选甲基、乙基、正丙基、异丙基、正丁基、叔丁基、异丁基或仲丁基；(22) In R 2-1-1 , the C 1 -C 6 alkyl group is a C 1 -C 4 alkyl group, preferably methyl, ethyl, n-propyl, isopropyl, n-butyl, tert-butyl, isobutyl or sec-butyl;
    (23)R2-1-2中，所述C1-C6的烷基为C1-C4的烷基，优选甲基、乙基、正丙基、异丙基、正丁基、叔丁基、异丁基或仲丁基；(23) In R 2-1-2 , the C 1 -C 6 alkyl group is a C 1 -C 4 alkyl group, preferably methyl, ethyl, n-propyl, isopropyl, n-butyl, tert-butyl, isobutyl or sec-butyl;
    (24)R2-1-3中，所述C1-C6的烷基为C1-C4的烷基，优选甲基、乙基、正丙基、异丙基、正丁基、叔丁基、异丁基或仲丁基；(24) In R 2-1-3 , the C 1 -C 6 alkyl group is a C 1 -C 4 alkyl group, preferably methyl, ethyl, n-propyl, isopropyl, n-butyl, tert-butyl, isobutyl or sec-butyl;
    (25)各个R2-1-4中，所述C1-C6的烷基为C1-C4的烷基，优选甲基、乙基、正丙基、异丙基、正丁基、叔丁基、异丁基或仲丁基；(25) In each R 2-1-4 , the C 1 -C 6 alkyl group is a C 1 -C 4 alkyl group, preferably methyl, ethyl, n-propyl, isopropyl, n-butyl , tert-butyl, isobutyl or sec-butyl;
    和(26)各个R2-2中，所述C1-C10的烷基为C1-C6的烷基，优选甲基、乙基、正丙基、异丙基、正丁基、叔丁基、异丁基或仲丁基。and (26) In each R 2-2 , the C 1 -C 10 alkyl group is a C 1 -C 6 alkyl group, preferably methyl, ethyl, n-propyl, isopropyl, n-butyl, tert-butyl, isobutyl or sec-butyl.
  3. 如权利要求1所述的如式I所示的含氮链状化合物或其药学上可接受的盐，其特征在于，The nitrogen-containing chain compound represented by formula I or a pharmaceutically acceptable salt thereof according to claim 1, characterized in that,
    所述如式I所示的含氮链状化合物为如式Ia、Ib、Ic、Id或Ie所示的化合物为 The nitrogen-containing chain compound represented by formula I is a compound represented by formula Ia, Ib, Ic, Id or Ie.
  4. 如权利要求1所述的如式I所示的含氮链状化合物或其药学上可接受的盐，其特征在于，所述如式I所示的含氮链状化合物为方案1、方案2、方案3、方案4或方案5中的任意一种：The nitrogen-containing chain compound represented by Formula I or a pharmaceutically acceptable salt thereof according to claim 1, characterized in that the nitrogen-containing chain compound represented by Formula I is Scheme 1 or Scheme 2. , any one of Option 3, Option 4 or Option 5:
    方案1、plan 1,
    X为C1-C4的亚烷基或C3-C6的亚环烷基；X is a C 1 -C 4 alkylene group or a C 3 -C 6 cycloalkylene group;
    Y为C1-C10的亚烷基；Y is a C 1 -C 10 alkylene group;
    Z为C1-C10的亚烷基；Z is a C 1 -C 10 alkylene group;
    A1和A2独立地为 A 1 and A 2 are independently
    R1为未取代的或被1个、2个或3个R1-1取代的C1-C20的烷基；R 1 is a C 1 -C 20 alkyl group that is unsubstituted or substituted by 1, 2 or 3 R 1-1 ;
    各个R1-1独立地为羟基、C1-C10的烷基或未取代的或被1个、2个或3个R1-1-4取代的C3-C15的环烷基；Each R 1-1 is independently a hydroxyl group, a C 1 -C 10 alkyl group or a C 3 -C 15 cycloalkyl group that is unsubstituted or substituted by 1, 2 or 3 R 1-1-4 ;
    各个R1-1-4独立地为C1-C6的烷基；Each R 1-1-4 is independently a C 1 -C 6 alkyl group;
    R2为未取代的或被1个、2个或3个R2-1取代的C1-C20的烷基；R 2 is a C 1 -C 20 alkyl group that is unsubstituted or substituted by 1, 2 or 3 R 2-1 ;
    各个R2-1独立地为羟基、C1-C10的烷基或未取代的或被1个、2个或3个R2-1-4取代的C3-C15的环烷基；Each R 2-1 is independently a hydroxyl group, a C 1 -C 10 alkyl group or a C 3 -C 15 cycloalkyl group that is unsubstituted or substituted by 1, 2 or 3 R 2-1-4 ;
    各个R2-1-4独立地为C1-C6的烷基；Each R 2-1-4 is independently a C 1 -C 6 alkyl group;
    方案2、Scenario 2,
    X为C1-C4的亚烷基或C3-C6的亚环烷基；X is a C 1 -C 4 alkylene group or a C 3 -C 6 cycloalkylene group;
    Y为C4-C8的亚烷基；Y is a C 4 -C 8 alkylene group;
    Z为C4-C8的亚烷基； Z is a C 4 -C 8 alkylene group;
    A1为其中a与R1相连，b与Y相连；A 1 is Where a is connected to R 1 , b is connected to Y;
    A2为其中a与R2相连，b与Z相连；A 2 is Among them, a is connected to R 2 , and b is connected to Z;
    R1为未取代的或被1个、2个或3个R1-1取代的C1-C20的烷基；R 1 is a C 1 -C 20 alkyl group that is unsubstituted or substituted by 1, 2 or 3 R 1-1 ;
    各个R1-1独立地为羟基、C1-C6的烷基或未取代的或被1个、2个或3个R1-1-4取代的C3-C6的环烷基；Each R 1-1 is independently a hydroxyl group, a C 1 -C 6 alkyl group or a C 3 -C 6 cycloalkyl group that is unsubstituted or substituted by 1, 2 or 3 R 1-1-4 ;
    各个R1-1-4独立地为C1-C6的烷基；Each R 1-1-4 is independently a C 1 -C 6 alkyl group;
    R2为未取代的或被1个、2个或3个R2-1取代的C1-C20的烷基；R 2 is a C 1 -C 20 alkyl group that is unsubstituted or substituted by 1, 2 or 3 R 2-1 ;
    各个R2-1独立地为羟基、C1-C6的烷基或未取代的或被1个、2个或3个R2-1-4取代的C3-C6的环烷基；Each R 2-1 is independently a hydroxyl group, a C 1 -C 6 alkyl group or a C 3 -C 6 cycloalkyl group that is unsubstituted or substituted by 1, 2 or 3 R 2-1-4 ;
    各个R2-1-4独立地为C1-C6的烷基；Each R 2-1-4 is independently a C 1 -C 6 alkyl group;
    当X为C1-C4的亚烷基时，R1为被1个、2个或3个羟基取代的C1-C20的烷基、未取代的或被1个、2个或3个R1-1-4取代的环戊烷、 When X is a C 1 -C 4 alkylene group, R 1 is a C 1 -C 20 alkyl group substituted by 1, 2 or 3 hydroxyl groups, unsubstituted, or R 1-1-4 substituted cyclopentane,
    方案3、Option 3.
    X为C1-C4的亚烷基；X is a C 1 -C 4 alkylene group;
    Y为C4-C8的亚烷基；Y is a C 4 -C 8 alkylene group;
    Z为C4-C8的亚烷基；Z is a C 4 -C 8 alkylene group;
    A1为其中a与R1相连，b与Y相连；A 1 is Where a is connected to R 1 , b is connected to Y;
    A2为其中a与R2相连，b与Z相连；A 2 is Among them, a is connected to R 2 , and b is connected to Z;
    R1为被1个、2个或3个羟基或环戊基取代的C1-C15的烷基、 R 1 is a C 1 -C 15 alkyl group substituted by 1, 2 or 3 hydroxyl groups or cyclopentyl groups,
    R2为被1个、2个或3个R2-1取代的C1-C15的烷基； R 2 is a C 1 -C 15 alkyl group substituted by 1, 2 or 3 R 2-1 ;
    各个R2-1独立地为C1-C10的烷基；Each R 2-1 is independently a C 1 -C 10 alkyl group;
    方案4、Option 4.
    X为C1-C4的亚烷基或C3-C6的亚环烷基；X is a C 1 -C 4 alkylene group or a C 3 -C 6 cycloalkylene group;
    Y为C4-C8的亚烷基；Y is a C 4 -C 8 alkylene group;
    Z为C4-C8的亚烷基；Z is a C 4 -C 8 alkylene group;
    A1为其中a与R1相连，b与Y相连；A 1 is Where a is connected to R 1 , b is connected to Y;
    A2为其中a与R2相连，b与Z相连；A 2 is Among them, a is connected to R 2 , and b is connected to Z;
    R1为未取代的或被1个、2个或3个R1-1取代的C1-C20的烷基；R 1 is a C 1 -C 20 alkyl group that is unsubstituted or substituted by 1, 2 or 3 R 1-1 ;
    各个R1-1独立地为羟基、C1-C6的烷基或未取代的或被1个、2个或3个R1-1-4取代的C3-C6的环烷基；Each R 1-1 is independently a hydroxyl group, a C 1 -C 6 alkyl group or a C 3 -C 6 cycloalkyl group that is unsubstituted or substituted by 1, 2 or 3 R 1-1-4 ;
    各个R1-1-4独立地为C1-C6的烷基；Each R 1-1-4 is independently a C 1 -C 6 alkyl group;
    R2为未取代的或被1个、2个或3个R2-1取代的C1-C20的烷基；R 2 is a C 1 -C 20 alkyl group that is unsubstituted or substituted by 1, 2 or 3 R 2-1 ;
    各个R2-1独立地为羟基、C1-C8的烷基或未取代的或被1个、2个或3个R2-1-4取代的C3-C6的环烷基；Each R 2-1 is independently a hydroxyl group, a C 1 -C 8 alkyl group or a C 3 -C 6 cycloalkyl group that is unsubstituted or substituted by 1, 2 or 3 R 2-1-4 ;
    各个R2-1-4独立地为C1-C6的烷基；Each R 2-1-4 is independently a C 1 -C 6 alkyl group;
    当X为C1-C4的亚烷基时，R1为被1个、2个或3个羟基取代的C1-C20的烷基、未取代的或被1个、2个或3个R1-1-4取代的环戊烷、 When X is a C 1 -C 4 alkylene group, R 1 is a C 1 -C 20 alkyl group substituted by 1, 2 or 3 hydroxyl groups, unsubstituted, or R 1-1-4 substituted cyclopentane,
    方案5、Option 5.
    X为C1-C4的亚烷基；X is a C 1 -C 4 alkylene group;
    Y为C4-C8的亚烷基；Y is a C 4 -C 8 alkylene group;
    Z为C4-C8的亚烷基；Z is a C 4 -C 8 alkylene group;
    A1为其中a与R1相连，b与Y相连； A 1 is Where a is connected to R 1 , b is connected to Y;
    A2为其中a与R2相连，b与Z相连；A 2 is Among them, a is connected to R 2 , and b is connected to Z;
    R1为被1个、2个或3个羟基或环戊烷取代的C1-C15的烷基、 R 1 is a C 1 -C 15 alkyl group substituted by 1, 2 or 3 hydroxyl groups or cyclopentane,
    R2为被1个、2个或3个R2-1取代的C1-C15的烷基；R 2 is a C 1 -C 15 alkyl group substituted by 1, 2 or 3 R 2-1 ;
    各个R2-1独立地为C1-C10的烷基或未取代的或被1个、2个或3个R1-1-4取代的C3-C6的环烷基；Each R 2-1 is independently a C 1 -C 10 alkyl group or a C 3 -C 6 cycloalkyl group that is unsubstituted or substituted by 1, 2 or 3 R 1-1-4 ;
    各个R2-1-4独立地为C1-C6的烷基。Each R 2-1-4 is independently a C 1 -C 6 alkyl group.
  5. 如权利要求1所述的如式I所示的含氮链状化合物或其药学上可接受的盐，其特征在于，所述如式I所示的含氮链状化合物满足如下条件中的一种或多种：The nitrogen-containing chain compound represented by formula I or a pharmaceutically acceptable salt thereof according to claim 1, characterized in that the nitrogen-containing chain compound represented by formula I satisfies one of the following conditions: One or more species:
    (1)X为C1-C4的亚烷基或C3-C6的亚环烷基；(1) X is a C 1 -C 4 alkylene group or a C 3 -C 6 cycloalkylene group;
    (2)Y为C1-C10的亚烷基；(2) Y is a C 1 -C 10 alkylene group;
    (3)Z为C1-C10的亚烷基；(3) Z is a C 1 -C 10 alkylene group;
    (4)R1为未取代的或被1个、2个或3个R1-1取代的C1-C20的烷基；(4) R 1 is an unsubstituted or C 1 -C 20 alkyl group substituted by 1, 2 or 3 R 1-1 ;
    (5)各个R1-1独立地为羟基、C1-C10的烷基或未取代的或被1个、2个或3个R1-1-4取代的C3-C15的环烷基；(5) Each R 1-1 is independently a hydroxyl group, a C 1 -C 10 alkyl group, or a C 3 -C 15 ring that is unsubstituted or substituted by 1, 2 or 3 R 1-1-4 alkyl;
    (6)R2为未取代的或被1个、2个或3个R2-1取代的C1-C20的烷基；(6) R 2 is a C 1 -C 20 alkyl group that is unsubstituted or substituted by 1, 2 or 3 R 2-1 ;
    (7)各个R2-1独立地为羟基、C1-C10的烷基或未取代的或被1个、2个或3个R2-1-4取代的C3-C15的环烷基；(7) Each R 2-1 is independently a hydroxyl group, a C 1 -C 10 alkyl group, or a C 3 -C 15 ring that is unsubstituted or substituted by 1, 2 or 3 R 2-1-4 alkyl;
    和(8)R1与R2相同。Same as (8) R 1 and R 2 .
  6. 如权利要求5所述的如式I所示的含氮链状化合物或其药学上可接受的盐，其特征在于，所述如式I所示的含氮链状化合物满足如下条件中的一种或多种：The nitrogen-containing chain compound represented by formula I or a pharmaceutically acceptable salt thereof according to claim 5, characterized in that the nitrogen-containing chain compound represented by formula I satisfies one of the following conditions: One or more species:
    (1)Y为C4-C8的亚烷基；(1) Y is a C 4 -C 8 alkylene group;
    (2)Z为C4-C8的亚烷基；(2) Z is a C 4 -C 8 alkylene group;
    (3)各个R1-1独立地为羟基、C1-C6的烷基或未取代的或被1个、2个或3个R1-1-4取代的C3-C6的环烷基；(3) Each R 1-1 is independently a hydroxyl group, a C 1 -C 6 alkyl group, or a C 3 -C 6 ring that is unsubstituted or substituted by 1, 2 or 3 R 1-1-4 alkyl;
    (4)各个R2-1独立地为羟基、C1-C6的烷基或未取代的或被1个、2个或3个R2-1-4取代的C3-C6的环烷基；(4) Each R 2-1 is independently a hydroxyl group, a C 1 -C 6 alkyl group, or a C 3 -C 6 ring that is unsubstituted or substituted by 1, 2 or 3 R 2-1-4 alkyl;
    和(5)当X为C1-C4的亚烷基时，R1为被1个、2个或3个羟基取代的C1-C20的烷基、未取代 的或被1个、2个或3个R1-1-4取代的环戊烷、 and (5) when X is a C 1 -C 4 alkylene group, R 1 is a C 1 -C 20 alkyl group substituted by 1, 2 or 3 hydroxyl groups, unsubstituted or cyclopentane substituted by 1, 2 or 3 R 1-1-4 ,
  7. 如权利要求6所述的如式I所示的含氮链状化合物或其药学上可接受的盐，其特征在于，所述如式I所示的含氮链状化合物满足如下条件中的一种或多种：The nitrogen-containing chain compound represented by formula I or a pharmaceutically acceptable salt thereof according to claim 6, characterized in that the nitrogen-containing chain compound represented by formula I satisfies one of the following conditions: One or more species:
    (1)X为 (1)X is
    (2)Y为 (2) Y is
    (3)Z为 (3) Z is
    (4)R1为 (4)R 1 is
    和(5)R2为 and (5)R 2 is
  8. 如权利要求1所述的如式I所示的含氮链状化合物或其药学上可接受的盐，其特征在于，所述如式I所示的含氮链状化合物为如下任一化合物：











    The nitrogen-containing chain compound represented by formula I or a pharmaceutically acceptable salt thereof according to claim 1, characterized in that the nitrogen-containing chain compound represented by formula I is any of the following compounds:











  9. 一种如式I所示的含氮链状化合物的制备方法，其特征在于，其包括如下步骤：溶剂中，在碱和碘盐的存在下，如式I-1所示的化合物与如式I-2所示的化合物进行如下式所示的偶联反应，即可；
    A method for preparing a nitrogen-containing chain compound represented by formula I, characterized in that it includes the following steps: in a solvent, in the presence of a base and an iodide salt, a compound represented by formula I-1 is mixed with a compound represented by formula I The compound represented by I-2 can undergo a coupling reaction as shown in the following formula;
    M为卤素，X、Y、Z、A1、A2、R1和R2如权利要求1-8任一项所述；M is halogen, and X, Y, Z, A 1 , A 2 , R 1 and R 2 are as described in any one of claims 1-8;
    较佳地，所述如式I所示的含氮链状化合物的制备方法满足如下条件中的一种或多种：Preferably, the preparation method of the nitrogen-containing chain compound shown in Formula I satisfies one or more of the following conditions:
    (1)所述偶联反应中，所述卤素为氟、氯、溴或碘，例如溴；(1) In the coupling reaction, the halogen is fluorine, chlorine, bromine or iodine, such as bromine;
    (2)所述偶联反应中，所述如式I-1所示的化合物与所述如式I-2所示的化合物的摩尔比为1:(1-2)，例如1:1.2；(2) In the coupling reaction, the molar ratio of the compound represented by Formula I-1 to the compound represented by Formula I-2 is 1:(1-2), such as 1:1.2;
    (3)所述偶联反应中，所述碱为碱式碳酸盐，例如K2CO3；(3) In the coupling reaction, the base is a basic carbonate, such as K 2 CO 3 ;
    (4)所述偶联反应中，所述如式I-1所示的化合物与所述碱的摩尔比为1:(1-5)，例如1:3.5；(4) In the coupling reaction, the molar ratio of the compound represented by Formula I-1 to the base is 1:(1-5), such as 1:3.5;
    (5)所述偶联反应中，所述碘盐为碱式碘盐，例如KI；(5) In the coupling reaction, the iodide salt is a basic iodide salt, such as KI;
    (6)所述偶联反应中，所述如式I-1所示的化合物与所述碘盐的摩尔比为1:(1-2)，例如1:1.2；(6) In the coupling reaction, the molar ratio of the compound represented by Formula I-1 to the iodide salt is 1:(1-2), such as 1:1.2;
    (7)所述偶联反应中，所述溶剂为醚类溶剂或/和腈类溶剂；所述醚类溶剂可为甲基叔丁基醚；所述腈类溶剂可为乙腈；所述腈类溶剂与醚类溶剂的体积比可为1:1；(7) In the coupling reaction, the solvent is an ether solvent or/and a nitrile solvent; the ether solvent can be methyl tert-butyl ether; the nitrile solvent can be acetonitrile; the nitrile The volume ratio of solvent-like solvents to ether-like solvents can be 1:1;
    (8)所述偶联反应中，所述如式I-1所示的化合物与所述溶剂的质量体积比为10mg/mL-65mg/mL， 例如15mg/mL、25mg/mL、40mg/mL或58.5mg/mL；(8) In the coupling reaction, the mass-to-volume ratio of the compound represented by Formula I-1 to the solvent is 10 mg/mL-65 mg/mL, For example, 15mg/mL, 25mg/mL, 40mg/mL or 58.5mg/mL;
    和(9)所述偶联反应中，所述偶联反应的反应温度为70℃-90℃，例如80℃。In the coupling reaction described in (9), the reaction temperature of the coupling reaction is 70°C-90°C, such as 80°C.
  10. 一种脂质载体，其特征在于，其包括物质Z，所述物质Z为如权利要求1-8任一项所述的如式I所示化合物或其药学可接受的盐。A lipid carrier, characterized in that it includes substance Z, which is a compound represented by formula I according to any one of claims 1 to 8 or a pharmaceutically acceptable salt thereof.
  11. 如权利要求10所述的脂质载体，其特征在于，所述脂质载体满足如下条件中的一种或多种：The lipid carrier according to claim 10, characterized in that the lipid carrier satisfies one or more of the following conditions:
    (1)所述脂质载体还包括稀释剂；(1) The lipid carrier also includes a diluent;
    (2)所述脂质载体还包括磷脂；(2) The lipid carrier also includes phospholipids;
    (3)所述脂质载体还包括PEG脂质；(3) The lipid carrier also includes PEG lipid;
    和(4)所述脂质载体还包括甾醇。and (4) the lipid carrier further includes sterols.
  12. 如权利要求11所述的脂质载体，其特征在于，所述脂质载体满足如下条件中的一种或多种：The lipid carrier according to claim 11, characterized in that the lipid carrier satisfies one or more of the following conditions:
    (1)所述稀释剂为磷酸盐缓冲液或Tris缓冲液；(1) The diluent is phosphate buffer or Tris buffer;
    (2)所述磷脂为具有带电极性端和脂肪链非极性端的磷脂类分子，例如二硬脂酰基磷脂酰胆碱、二肉豆蔻酰磷酸胆碱、二油酰磷酸胆碱、棕榈酰磷酸胆碱、1,2-二硬脂酰磷酸胆碱、二十一烷酰磷酸胆碱或棕榈酰磷酸胆碱；(2) The phospholipid is a phospholipid molecule with a charged polar end and a non-polar end of the fatty chain, such as distearoyl phosphatidylcholine, dimyristoyl phosphocholine, dioleoyl phosphocholine, palmitoyl Phosphocholine, 1,2-distearyl phosphocholine, icosanoyl phosphocholine, or palmitoyl phosphocholine;
    (3)所述PEG脂质为具有聚乙二醇亲水端修饰的脂质分子；较佳地，所述PEG脂质优选选自PEG修饰的磷脂酰乙醇胺、PEG修饰的磷脂酸、PEG修饰的神经酰胺、PEG修饰的二烷基胺、PEG修饰的二酰基甘油和PEG修饰的二烷基甘油中的一种或多种，例如所述PEG脂质为具PEG修饰的二肉豆蔻酰甘油；(3) The PEG lipid is a lipid molecule modified with a polyethylene glycol hydrophilic end; preferably, the PEG lipid is preferably selected from the group consisting of PEG-modified phosphatidylethanolamine, PEG-modified phosphatidic acid, and PEG-modified One or more of ceramide, PEG-modified dialkylamine, PEG-modified diacylglycerol and PEG-modified dialkylglycerol, for example, the PEG lipid is PEG-modified dimyristoylglycerol ;
    (4)所述甾醇包括动物性、植物性或菌类甾醇；较佳地，所述甾醇选自胆固醇、谷甾醇、麦角甾醇、菜油甾醇、豆甾醇、芸苔甾醇、番茄碱、熊果酸和α-生育酚中的一种或多种，例如胆固醇；(4) The sterols include animal, plant or fungal sterols; preferably, the sterols are selected from cholesterol, sitosterol, ergosterol, campesterol, stigmasterol, brassicosterol, tomatine, and ursolic acid and one or more of alpha-tocopherols, such as cholesterol;
    (5)所述脂质载体中，所述物质Z与甾醇的摩尔比为0.5～5:1，优选为0.5～3:1，例如0.6～2:1；(5) In the lipid carrier, the molar ratio of the substance Z to the sterol is 0.5 to 5:1, preferably 0.5 to 3:1, such as 0.6 to 2:1;
    (6)所述脂质载体中，所述物质Z与磷脂的摩尔比为1～15:1，优选为2～8:1，例如3～6:1；(6) In the lipid carrier, the molar ratio of the substance Z to the phospholipid is 1 to 15:1, preferably 2 to 8:1, such as 3 to 6:1;
    (7)所述脂质载体中，所述物质Z与PEG脂质的摩尔比为20～130:1，优选为20～80:1，例如20～40:1；(7) In the lipid carrier, the molar ratio of the substance Z to the PEG lipid is 20 to 130:1, preferably 20 to 80:1, such as 20 to 40:1;
    (8)所述物质Z的摩尔含量为30mol％至60mol％；(8) The molar content of substance Z is 30 mol% to 60 mol%;
    (9)所述磷脂的摩尔含量为0mol％至30mol％；(9) The molar content of the phospholipid is 0 mol% to 30 mol%;
    (10)所述甾醇的摩尔含量为15mol％至55mol％；(10) The molar content of the sterol is 15 mol% to 55 mol%;
    和(11)所述PEG脂质的摩尔含量为0mol％至10mol％；and (11) the molar content of the PEG lipid is 0 mol% to 10 mol%;
    较佳地，所述脂质载体由所述物质Z、所述稀释剂、所述磷脂、所述PEG脂质和所述甾醇组成。Preferably, the lipid carrier is composed of the substance Z, the diluent, the phospholipid, the PEG lipid and the sterol.
  13. 一种脂质纳米颗粒，其特征在于，其包括治疗剂和/或预防剂以及如权利要求10-12任一项所述的脂质载体。A lipid nanoparticle, characterized in that it includes a therapeutic agent and/or a preventive agent and the lipid carrier according to any one of claims 10-12.
  14. 如权利要求13所述的脂质纳米颗粒，其特征在于，所述脂质纳米颗粒满足如下条件中的一种或多种：The lipid nanoparticle according to claim 13, wherein the lipid nanoparticle satisfies one or more of the following conditions:
    (1)所述治疗剂和/或预防剂为一种或两种及以上核酸；较佳地，所述治疗剂和/或预防剂为单链 脱氧核糖核酸、双链DNA、小干扰RNA、不对称双链小干扰RNA、微小RNA、小发夹RNA、环状RNA、转运RNA或信使RNA，优选为mRNA，例如萤火虫荧光素酶mRNA或SARS-CoV-2刺突蛋白mRNA；(1) The therapeutic agent and/or preventive agent is one or two or more nucleic acids; preferably, the therapeutic agent and/or preventive agent is single-stranded Deoxyribonucleic acid, double-stranded DNA, small interfering RNA, asymmetric double-stranded small interfering RNA, microRNA, small hairpin RNA, circular RNA, transfer RNA or messenger RNA, preferably mRNA, such as firefly luciferase mRNA or SARS -CoV-2 spike protein mRNA;
    (2)所述脂质纳米颗粒中氮磷比为2:1～30:1，优选为2:1～20:1，例如3:1～20:1，还例如3:1～16:1；(2) The nitrogen to phosphorus ratio in the lipid nanoparticles is 2:1 to 30:1, preferably 2:1 to 20:1, such as 3:1 to 20:1, or 3:1 to 16:1. ;
    (3)所述脂质纳米颗粒中，所述脂质载体与治疗剂和/或预防剂的质量比为3～80:1，优选为6～60:1(3) In the lipid nanoparticles, the mass ratio of the lipid carrier to the therapeutic agent and/or preventive agent is 3 to 80:1, preferably 6 to 60:1.
    (4)所述脂质纳米颗粒的粒径为10～250nm，例如为40～250nm，进一步为50～250nm，或40～150nm，或60～150nm；(4) The particle size of the lipid nanoparticles is 10-250nm, for example, 40-250nm, further 50-250nm, or 40-150nm, or 60-150nm;
    和(5)所述脂质纳米颗粒中，所述脂质载体包裹所述治疗剂和/或预防剂。and (5) in the lipid nanoparticles, the lipid carrier encapsulates the therapeutic agent and/or preventive agent.
  15. 一种组合物，其特征在于，其包括物质Z，所述物质Z为如权利要求1-8任一项所述的如式I所示化合物或其药学可接受的盐。A composition, characterized in that it includes substance Z, which is a compound represented by formula I according to any one of claims 1 to 8 or a pharmaceutically acceptable salt thereof.
  16. 如权利要求15所述的组合物，其特征在于，所述组合物还包括稀释剂、磷脂、PEG脂质、甾醇和治疗剂和/或预防剂中的一种或多种；The composition of claim 15, wherein the composition further includes one or more of a diluent, a phospholipid, a PEG lipid, a sterol, and a therapeutic and/or preventive agent;
    较佳地，所述组合物中，所述稀释剂、磷脂、PEG脂质和甾醇如权利要求12所述；和/或，所述治疗剂和/或预防剂如权利要求14所述；Preferably, in the composition, the diluent, phospholipid, PEG lipid and sterol are as described in claim 12; and/or the therapeutic agent and/or preventive agent is as described in claim 14;
    更佳地，所述组合物中，所述物质Z与所述稀释剂、磷脂、PEG脂质和甾醇中的一种或多种形成如权利要求10-12任一项所述的脂质载体；和或，所述组合物中，所述治疗剂和/或预防剂的包封率为至少50％，优选为至少为70％；More preferably, in the composition, the substance Z and one or more of the diluent, phospholipid, PEG lipid and sterol form a lipid carrier as claimed in any one of claims 10-12 ; And or, in the composition, the encapsulation rate of the therapeutic agent and/or preventive agent is at least 50%, preferably at least 70%;
    还佳地，所述组合物中，所述脂质载体与所述治疗剂和/或预防剂形成如权利要求13或14所述的脂质纳米颗粒；和或，所述组合物中，所述组合物的多分散指数为不高于0.5，例如不高于0.3。 Preferably, in the composition, the lipid carrier and the therapeutic agent and/or preventive agent form lipid nanoparticles as claimed in claim 13 or 14; and or, in the composition, the lipid nanoparticles The composition has a polydispersity index of not higher than 0.5, for example not higher than 0.3.
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CN113636947A (en) * 2015-10-28 2021-11-12 爱康泰生治疗公司 Novel lipid and lipid nanoparticle formulations for delivery of nucleic acids
CN114044741A (en) * 2022-01-13 2022-02-15 北京悦康科创医药科技股份有限公司 Cationic lipid compound, composition containing same and application
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CN113636947A (en) * 2015-10-28 2021-11-12 爱康泰生治疗公司 Novel lipid and lipid nanoparticle formulations for delivery of nucleic acids
CN114044741A (en) * 2022-01-13 2022-02-15 北京悦康科创医药科技股份有限公司 Cationic lipid compound, composition containing same and application
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