WO2023024515A1 - Novel cationic lipid compound - Google Patents

Novel cationic lipid compound Download PDF

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Publication number
WO2023024515A1
WO2023024515A1 PCT/CN2022/084347 CN2022084347W WO2023024515A1 WO 2023024515 A1 WO2023024515 A1 WO 2023024515A1 CN 2022084347 W CN2022084347 W CN 2022084347W WO 2023024515 A1 WO2023024515 A1 WO 2023024515A1
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compound
dmso
nmr
composition
synthesis
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PCT/CN2022/084347
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French (fr)
Chinese (zh)
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黄才古
徐天祥
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广州谷森制药有限公司
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Publication of WO2023024515A1 publication Critical patent/WO2023024515A1/en

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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
    • A61K47/22—Heterocyclic compounds, e.g. ascorbic acid, tocopherol or pyrrolidones
    • 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
    • A61K47/24—Organic compounds, e.g. natural or synthetic hydrocarbons, polyolefins, mineral oil, petrolatum or ozokerite containing atoms other than carbon, hydrogen, oxygen, halogen, nitrogen or sulfur, e.g. cyclomethicone or phospholipids
    • 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
    • A61K47/28—Steroids, e.g. cholesterol, bile acids or glycyrrhetinic acid
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K48/00—Medicinal preparations containing genetic material which is inserted into cells of the living body to treat genetic diseases; Gene therapy
    • 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
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07B—GENERAL METHODS OF ORGANIC CHEMISTRY; APPARATUS THEREFOR
    • C07B59/00—Introduction of isotopes of elements into organic compounds ; Labelled organic compounds per se
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D295/00—Heterocyclic compounds containing polymethylene-imine rings with at least five ring members, 3-azabicyclo [3.2.2] nonane, piperazine, morpholine or thiomorpholine rings, having only hydrogen atoms directly attached to the ring carbon atoms
    • C07D295/04—Heterocyclic compounds containing polymethylene-imine rings with at least five ring members, 3-azabicyclo [3.2.2] nonane, piperazine, morpholine or thiomorpholine rings, having only hydrogen atoms directly attached to the ring carbon atoms with substituted hydrocarbon radicals attached to ring nitrogen atoms
    • C07D295/12—Heterocyclic compounds containing polymethylene-imine rings with at least five ring members, 3-azabicyclo [3.2.2] nonane, piperazine, morpholine or thiomorpholine rings, having only hydrogen atoms directly attached to the ring carbon atoms with substituted hydrocarbon radicals attached to ring nitrogen atoms substituted by singly or doubly bound nitrogen atoms
    • C07D295/125—Heterocyclic compounds containing polymethylene-imine rings with at least five ring members, 3-azabicyclo [3.2.2] nonane, piperazine, morpholine or thiomorpholine rings, having only hydrogen atoms directly attached to the ring carbon atoms with substituted hydrocarbon radicals attached to ring nitrogen atoms substituted by singly or doubly bound nitrogen atoms with the ring nitrogen atoms and the substituent nitrogen atoms attached to the same carbon chain, which is not interrupted by carbocyclic rings
    • C07D295/13—Heterocyclic compounds containing polymethylene-imine rings with at least five ring members, 3-azabicyclo [3.2.2] nonane, piperazine, morpholine or thiomorpholine rings, having only hydrogen atoms directly attached to the ring carbon atoms with substituted hydrocarbon radicals attached to ring nitrogen atoms substituted by singly or doubly bound nitrogen atoms with the ring nitrogen atoms and the substituent nitrogen atoms attached to the same carbon chain, which is not interrupted by carbocyclic rings to an acyclic saturated chain

Definitions

  • the present invention provides novel cationic lipids that can be used in combination with other lipid components (such as neutral lipids, steroids, and polymer-conjugated lipids) to form a nucleic acid mRNA lipid nanoparticle composition combining a A method of delivering one or more therapeutic and/or prophylactic agents to a mammalian cell or organ and/or producing a polypeptide in a mammalian cell or organ.
  • lipid components such as neutral lipids, steroids, and polymer-conjugated lipids
  • the lipid nanoparticle compositions of the invention may also include, in specified proportions, one or more cationic and/or ionizable amino lipids, neutral lipids including polyunsaturated lipids, Lipids, polymer-conjugated lipids, steroids, and/or therapeutic and/or prophylactic agents.
  • nucleic acids Efficient targeted delivery of bioactive substances such as small molecule drugs, proteins and nucleic acids presents a persistent medical problem. Specifically, delivery of nucleic acids to cells is made difficult by the relative instability and low cell permeability of these species. Accordingly, there is a need to develop methods and compositions that facilitate the delivery of therapeutic and/or prophylactic agents, such as nucleic acids, to cells.
  • compositions generally comprise one or more "cationic" lipids, neutral lipids including polyunsaturated lipids (such as phospholipids), structured lipids (such as steroids) and/or polyethylene glycol-containing Lipids of alcohols (polymer-conjugated lipids).
  • Cationic lipids include, for example, amine-containing lipids that can be readily protonated.
  • RNA degradation in plasma has been used to prevent RNA degradation in plasma and promote Cellular uptake of oligonucleotides.
  • the improved lipid nanoparticles would provide optimized drug delivery, protect nucleic acids from degradation and clearance in serum, be suitable for systemic or local delivery, and provide intracellular delivery of nucleic acids.
  • these preferred lipid-nucleic acid particles should be well tolerated and provide a sufficient therapeutic index such that treatment of a patient at an effective dose of the nucleic acid does not create unacceptable toxicity and/or risk to the patient.
  • the present invention provides these and related advantages.
  • the present invention provides the following novel compounds and methods involving these compounds:
  • the present invention relates to compounds of the following structural formula (I):
  • R 1 , R 2 , R 3 , and R 4 in the structural formula "I" are two independent “hydrogen” isotopes (including isotopes
  • R and R are independently selected from any group consisting of "HH", “HD” and “DD”;
  • R3 and R4 are independently selected from any group consisting of "HH, HD”, “HD, HH”, “HH, DD”, “DD, HH”, “HD, DD” and “DD, DD”;
  • R 1 , R 2 , R 3 , and R 4 are all independent, but cannot be "HH” at the same time, that is, contain at least one "HD";
  • the compound has one of the structures shown in Table 1 below
  • composition comprising any one or more of the compounds of structural formula (I) and a therapeutic and/or prophylactic agent.
  • compositions comprising any one or more of the compounds of structure (I) and a therapeutic and/or prophylactic agent.
  • the composition comprises any of the compounds of structure (I) and a therapeutic and/or prophylactic agent and one or more compounds selected from the group consisting of neutral lipids, steroids, and polymer conjugates. Lipid excipients. Other pharmaceutically acceptable excipients and/or carriers are also included in various embodiments of the compositions.
  • the neutral lipid is selected from 1,2-distearoyl-sn-glycero-3-phosphocholine (DSPC), 1,2-dipalmitoyl-sn-glycero-3-phosphocholine Alkaline (DPPC), 1,2-Dimyristyl-sn-glycero-phosphocholine (DMPC), 1,2-dioleoyl-sn-glycero-3-phosphocholine (DOPC), 1-palm Acyl-2-oleoyl-sn-glycero-3-phosphoethanolamine (POPC), 1,2-dioleoyl-sn-glycero-3-phosphoethanolamine (DOPE), sphingomyelin (SM) and mixtures thereof.
  • the preferred neutral lipid is 1,2-distearoyl-sn-glycero-3-phosphocholine (DSPC).
  • the steroid is selected from the group consisting of cholesterol, fecal sterol, sitosterol, ergosterol, campesterol, stigmasterol, brassicasterol, tomatidine, ursolic acid, alpha-tocopherol, and mixtures thereof .
  • the preferred steroid is cholesterol.
  • the pegylated lipid is 1,2-dimyristoyl-sn-glycerol methoxypolyethylene glycol (PEG-DMG)
  • the composition ratio ranges from about 10 to 60 mol% of the compound, about 0 to 30 mol% of neutral lipids, about 10 to 55 mol% of steroids, and about 0 to 10 mol% of polymer-conjugated lipids. quality.
  • the therapeutic and/or prophylactic agent comprises a nucleic acid.
  • the nucleic acid is RNA, which is selected from the following composition: siRNA, aiRNA, miRNA, dsRNA, shRNA, mRNA and mixtures thereof.
  • the RNA is selected from mRNA.
  • the present invention relates to methods of administering therapeutic and/or prophylactic agents to a subject in need thereof, the method comprising preparing or providing any of the compositions described above and administering the composition to the subject combination.
  • the compounds of the invention can be administered as a drug substance, or can be formulated as a pharmaceutical composition.
  • the pharmaceutical composition of the present invention comprises a compound of structure (I) and one or more pharmaceutically acceptable carriers, diluents or excipients.
  • Compounds of structure (I) are effective to form lipid nanoparticles and deliver therapeutic and/or prophylactic agents. Appropriate concentrations and dosages can be readily determined by those skilled in the art.
  • compositions of the present invention may be by any of the acceptable modes of administration for agents of similar utility.
  • the pharmaceutical composition of the present invention can be formulated into preparations in solid, semi-solid, liquid or gaseous form, such as tablets, capsules, powders, granules, ointments, solutions, suspensions, suppositories, injections, inhalants, gels, microspheres and aerosols.
  • Typical routes of administration of such pharmaceutical compositions include, but are not limited to, oral, topical, transdermal, inhalation, parenteral, sublingual, buccal, rectal, vaginal and intranasal routes.
  • parenteral as used herein includes subcutaneous injections, intravenous, intramuscular, intradermal, intrasternal injection or infusion techniques.
  • the pharmaceutical compositions of the invention are formulated so as to allow the active ingredient contained therein to be bioavailable upon administration of the composition to a subject.
  • Compositions to be administered to a subject or patient are in the form of one or more dosage units, where a tablet may be a single dosage unit and a container of the compound in aerosol form of the invention may hold multiple dosage units. Current methods for the preparation of such dosage forms are known, or will be apparent, to those skilled in the art.
  • the composition to be administered will contain a therapeutically effective amount of a compound of the invention, or a pharmaceutically acceptable salt thereof, for treating the associated disease or condition in accordance with the teachings of the invention.
  • compositions of the present invention may be in solid or liquid form.
  • the carrier is particulate, such that the composition is in tablet or powder form.
  • the carrier can be a liquid in which case the composition is an oral syrup or an injectable liquid or an aerosol suitable for administration by inhalation.
  • the pharmaceutical composition When intended for oral administration, the pharmaceutical composition is preferably in solid or liquid form, where forms considered solid or liquid herein include semi-solid, semi-liquid, suspension and gel forms.
  • the pharmaceutical compositions can be formulated in the form of powders, granules, compressed tablets, pills, capsules, chewing gums, flakes and the like.
  • Such solid compositions will generally contain one or more inert diluents or edible carriers.
  • binders such as gelatin, cellulose, etc.
  • excipients such as lactose, etc.
  • disintegrants such as alginic acid, etc.
  • lubricants such as magnesium stearate, etc.
  • Glidants such as silica gel, etc.
  • sweeteners such as sucrose or saccharin
  • flavoring agents such as mint
  • coloring agents such as sucrose or saccharin
  • liquid carriers other than materials of the above type, such as polyethylene glycol or oil.
  • compositions may be in liquid form, such as syrups, solutions, emulsions or suspensions.
  • Liquids can be used for oral administration or for injectable delivery, as two examples.
  • the compositions of Urim contain, in addition to the compounds of the present invention, one or more of sweetening agents, preservatives, dyeing/coloring agents and darkening agents.
  • one or more of surfactants, preservatives, wetting agents, dispersing agents, suspending agents, buffers, stabilizers and isotonic agents may be included.
  • the liquid pharmaceutical composition of the present invention may include one or more of the following adjuvants: sterile diluents, such as water for injection, saline solution, preferably physiological saline, Ringer's solution, isotonic sodium chloride; fixed oils such as synthetic mono- or diglycerides, polyethylene glycol, glycerol, propylene glycol, or other solvents that may be used as a solvent or suspending medium; antibacterial agents , such as methylparaben, etc.; antioxidants, such as ascorbic acid or sodium bisulfite; chelating agents, such as ethylenedipic acid; buffering agents, such as acetate, clematine, or phosphate; Reagents for tonicity, such as sodium chloride or glucose; reagents used as cryoprotectants, such as sucrose or trehalose.
  • the parenteral preparation can be enclosed in ampoules, disposable syringes or multiple dose via
  • compositions of the present invention may be intended for topical administration, in which case the carrier may suitably comprise a solution base, emulsion base, ointment base or gel base.
  • the base may contain one or more of petrolatum, lanolin, polyethylene glycols, beeswax, mineral oil, diluents such as water and alcohol, and emulsifiers and stabilizers.
  • Thickening agents may be present in pharmaceutical compositions for topical administration.
  • the composition may include a transdermal patch or iontophoretic device.
  • compositions of the invention may include various materials which modify the physical form of solid or liquid dosage units.
  • the composition may include materials which form a coating shell around the active ingredient.
  • Materials forming the coating shell are generally inert and can be, sugar, shellac, and other enteric coating agents.
  • the active ingredient may be enclosed in a gelatin capsule.
  • compositions of the invention in solid or liquid form may include agents that bind to and thereby facilitate delivery of the compounds of the invention.
  • Suitable agents that may act in this capacity include monoclonal or polyclonal antibodies or proteins.
  • compositions of the invention may consist of dosage units which can be administered as an aerosol.
  • aerosol is used to denote systems ranging from those of a colloidal nature to those consisting of pressurized packs. Delivery may be by liquefied or compressed gas, or by a suitable pump system which dispenses the active ingredient. Aerosols of the compounds of the invention may be delivered as single-phase, bi-phasic or triphasic systems to deliver the active ingredient. Aerosol delivery includes the necessary containers, activators, valves, sub-containers, etc., which together may form a kit. Preferred aerosols can be determined by those skilled in the art without undue experimentation.
  • compositions of the present invention can be prepared by methods well known in the field of pharmacy.
  • Pharmaceutical compositions intended to be administered by injection can be prepared by combining the lipid nanoparticles of the present invention with sterile distilled water or other carriers into a solution.
  • Surfactants can be added to promote the formation of a uniform solution or suspension.
  • Surfactants are compounds that interact non-covalently with the compounds of the invention in order to facilitate dissolution or uniform suspension of the compounds in aqueous delivery systems.
  • composition of the present invention is administered in a therapeutically effective amount, which will vary depending on a number of factors, including the activity of the particular therapeutic agent used; the metabolic stability and duration of action of the therapeutic agent; The patient's age, weight, general health, sex, and diet; mode and timing of administration; rate of excretion; drug combination; severity of the specific case, etc.
  • compositions of the invention may also be administered concurrently with, before or after administration of one or more other therapeutic agents.
  • combination therapy includes administration of a single pharmaceutical dosage formulation of a composition of the invention and one or more additional active agents, as well as administration of a composition of the invention and each active agent in its own separate pharmaceutical dosage formulation.
  • compositions of the invention and other active agents may be administered to a subject together in a single oral dosage composition such as a tablet or capsule, or each agent may be administered in separate oral dosage formulations.
  • the compound of the invention and the one or more additional active agents may be administered at substantially the same time, or sequentially at times staggered from each other; it is understood that combination therapy includes all such dosing regimens.
  • novel deuterated cationic lipid compounds have achieved more advantageous physical and chemical properties, including more suitable pKa and better chemical stability, and are used in mRNA nanoliposome compositions, which can realize anti-ion Nucleic acid-like drugs are more effectively combined and delivered, and at the same time, their chemical structure is more stable, which is convenient for synthesis and beneficial for development as pharmaceutical excipients.
  • functional groups of intermediate compounds may need to be protected by suitable protecting groups.
  • suitable protecting groups for hydroxy include trialkylsilyl or diarylalkylsilyl, tetrahydrofuranyl, benzyl, and the like.
  • Suitable protecting groups for amino include t-butoxycarbonyl, benzyloxycarbonyl and the like.
  • Suitable protecting groups for carboxylic acids include hydrocarbyl, aryl or arylhydrocarbyl esters. Protecting groups can be added or removed according to standard techniques, known to those skilled in the art and described herein.
  • all compounds of the present invention that exist in free base or free acid form can be converted to their pharmaceutically acceptable salts by treatment with an appropriate inorganic or organic base or acid according to methods known to those skilled in the art. Salts of compounds of the present invention may be converted to their free base or acid forms by standard techniques.
  • Compound 1 can be synthesized according to the representative route described in Example 1.
  • Compound 2 can be synthesized according to the representative route described in Example 1.
  • Compound 3 can be synthesized according to the representative route described in Example 1.
  • Compound 4 can be synthesized according to the representative route described in Example 1.
  • Embodiment 6 is a diagrammatic representation of Embodiment 6
  • Compound 5 can be synthesized according to the representative route described in Example 1.
  • Embodiment 7 is a diagrammatic representation of Embodiment 7:
  • Compound 6 can be synthesized according to the representative route described in Example 1.
  • Embodiment 8 is a diagrammatic representation of Embodiment 8
  • Compound 7 can be synthesized according to the representative route described in Example 1.
  • Embodiment 9 is a diagrammatic representation of Embodiment 9:
  • Compound 8 can be synthesized according to the representative route described in Example 1.
  • Compound 9 can be synthesized according to the representative route described in Example 1.
  • Compound 10 can be synthesized according to the representative route described in Example 1.
  • Compound 11 can be synthesized according to the representative route described in Example 1.
  • Compound 12 can be synthesized according to the representative route described in Example 1.
  • Compound 13 can be synthesized according to the representative route described in Example 1.
  • Compound 14 can be synthesized according to the representative route described in Example 1.
  • Compound 15 can be synthesized according to the representative route described in Example 1.
  • Compound 16 can be synthesized according to the representative route described in Example 1.
  • Compound 17 can be synthesized according to the representative route described in Example 1.
  • Compound 18 can be synthesized according to the representative route described in Example 1.
  • Compound 19 can be synthesized according to the representative route described in Example 1.
  • Compound 20 can be synthesized according to the representative route described in Example 1.
  • Compound 21 can be synthesized according to the representative route described in Example 1.
  • Compound 22 can be synthesized according to the representative route described in Example 1.
  • Compound 23 can be synthesized according to the representative route described in Example 1.
  • Compound 24 can be synthesized according to the representative route described in Example 1.
  • Compound 25 can be synthesized according to the representative route described in Example 1.
  • Compound 26 can be synthesized according to the representative route described in Example 1.
  • Compound 27 can be synthesized according to the representative route described in Example 1.
  • Compound 28 can be synthesized according to the representative route described in Example 1.
  • Compound 29 can be synthesized according to the representative route described in Example 1.
  • Compound 30 can be synthesized according to the representative route described in Example 1.
  • Compound 31 can be synthesized according to the representative route described in Example 1.
  • Compound 32 can be synthesized according to the representative route described in Example 1.
  • Compound 33 can be synthesized according to the representative route described in Example 1.
  • Compound 34 can be synthesized according to the representative route described in Example 1.
  • Compound 35 can be synthesized according to the representative route described in Example 1.
  • Compound 36 can be synthesized according to the representative route described in Example 1.
  • Compound 37 can be synthesized according to the representative route described in Example 1.
  • Compound 38 can be synthesized according to the representative route described in Example 1.
  • Compound 39 can be synthesized according to the representative route described in Example 1.
  • Compound 40 can be synthesized according to the representative route described in Example 1.
  • Compound 41 can be synthesized according to the representative route described in Example 1.
  • Compound 42 can be synthesized according to the representative route described in Example 1.
  • Compound 43 can be synthesized according to the representative route described in Example 1.
  • Compound 44 can be synthesized according to the representative route described in Example 1.
  • Compound 45 can be synthesized according to the representative route described in Example 1.
  • Compound 46 can be synthesized according to the representative route described in Example 1.
  • Compound 47 can be synthesized according to the representative route described in Example 1.
  • Compound 48 can be synthesized according to the representative route described in Example 1.
  • Compound 49 can be synthesized according to the representative route described in Example 1.
  • Compound 50 can be synthesized according to the representative route described in Example 1.
  • Compound 51 can be synthesized according to the representative route described in Example 1.
  • Compound 53 can be synthesized according to the representative route described in Example 1.
  • Compound 53 can be synthesized according to the representative route described in Example 1.
  • Compound 54 can be synthesized according to the representative route described in Example 1.
  • Compound 55 can be synthesized according to the representative route described in Example 1.
  • Compound 56 can be synthesized according to the representative route described in Example 1.
  • Compound 57 can be synthesized according to the representative route described in Example 1.
  • Compound 58 can be synthesized according to the representative route described in Example 1.
  • Compound 59 can be synthesized according to the representative route described in Example 1.
  • Compound 60 can be synthesized according to the representative route described in Example 1.
  • Compound 61 can be synthesized according to the representative route described in Example 1.
  • Compound 62 can be synthesized according to the representative route described in Example 1.
  • Compound 63 can be synthesized according to the representative route described in Example 1.
  • Compound 64 can be synthesized according to the representative route described in Example 1.
  • Compound 65 can be synthesized according to the representative route described in Example 1.
  • Compound 66 can be synthesized according to the representative route described in Example 1.
  • Compound 67 can be synthesized according to the representative route described in Example 1.
  • Compound 68 can be synthesized according to the representative route described in Example 1.
  • Compound 69 can be synthesized according to the representative route described in Example 1.
  • Compound 70 can be synthesized according to the representative route described in Example 1.
  • Compound 71 can be synthesized according to the representative route described in Example 1.
  • Compound 72 can be synthesized according to the representative route described in Example 1.
  • Compound 73 can be synthesized according to the representative route described in Example 1.
  • Compound 74 can be synthesized according to the representative route described in Example 1.
  • Compound 75 can be synthesized according to the representative route described in Example 1.
  • Compound 76 can be synthesized according to the representative route described in Example 1.
  • Compound 77 can be synthesized according to the representative route described in Example 1.
  • Compound 78 can be synthesized according to the representative route described in Example 1.
  • the particle size of the lipid nanoparticles is approximately 65-105 nm in diameter, and in some cases, approximately 75-100 nm in diameter.
  • liver tissue approximately 50 mg was cut for analysis in 2 mL FastPrep tubes (MP Biomedicals, Solon OH). 1/4" ceramic balls (MP Biomedicals) were added to each tube, and 500 ⁇ L of Glo Lysis Buffer-GLB (Promega, Madison WI) equilibrated to room temperature was added to the liver tissue.
  • the FastPrep24 instrument MP Biomedicals
  • the Liver tissue was homogenized at 2x6.0m/s for 15 s. The homogenate was incubated at room temperature for 5 min, then diluted 1:4 in GLB and evaluated using the SteadyGlo luciferase assay system (Promega).
  • FLuc mRNA (L-6107) from Trilink Biotechnologies will express the luciferase protein, which was originally isolated from the firefly (Photinus pyralis). Fluc is commonly used in mammalian cell culture to measure gene expression and cell viability. It emits bioluminescence in the presence of the substrate luciferin. This capped and polyadenylated mRNA is completely replaced by 5-methylcytidine and pseudouridine.
  • the pKa of the formulated cationic lipid correlates with the effectiveness of the LNP for delivery of nucleic acids.
  • the preferred pKa range is 5-7.
  • the pKa of each cationic lipid was determined in lipid nanoparticles using an assay based on the fluorescence of 2-(p-toluidino)-6-phthanesulfonic acid (TNS).
  • TNS 2-(p-toluidino)-6-phthanesulfonic acid
  • the vesicles were diluted to contain 24uM lipid in 2mL buffer solution containing 10mM HEPES, 10mM MES, 10mM sodium acetate, 130mM NaCl, wherein the pH value was 2.5-11. Aliquots of TNS solution were added to give a final concentration of 1 uM, and after vortex mixing, fluorescence intensity was measured in a SLM Aminco Series 2 Luminescence Spectrophotometer at room temperature using excitation and emission wavelengths of 321 nm and 445 nm. Sigmoid best-fit analysis was applied to the fluorescence data, and pKa was measured as the pH that yielded half the maximum fluorescence intensity.
  • lipid nanoparticles containing FLuc mRNA were also used to formulate lipid nanoparticles containing FLuc mRNA (L-6107) using the ordered mixing method as described in Example 64.
  • Lipid nanoparticles were formulated using the following molar ratios: 50% cationic lipid/10% distearoylphosphatidylcholine (DSPC)/38% cholesterol/2% PEG lipid ("PEG-DMG", i.e., ( 1-(monomethoxy-polyethylene glycol)-2,3-dimyristoylglycerol, the average PEG molecular weight is 2000).
  • PEG-DMG i.e., ( 1-(monomethoxy-polyethylene glycol)-2,3-dimyristoylglycerol, the average PEG molecular weight is 2000.
  • Relative activity was determined by measuring luciferase expression in the liver. The activity was compared at doses of 0.3 and 1.0 mg mRNA/kg and expressed as ng luci

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Abstract

The present invention provides a lipid compound which can be used alone or in combination with other lipid components such as neutral lipids, charged lipids, steroids and/or analogs thereof, and/or polymer-conjugated lipids to form lipid nanoparticles for delivery of therapeutic and/or prophylactic agents. In some examples, lipid nanoparticles are used for delivering nucleic acids such as messenger RNAs and/or antisense RNAs. Also provided are a method for using such lipid nanoparticles to treat and/or prevent various diseases. In one embodiment, a compound having the structure of formula (I) below, or a salt, an isomer, or an N-oxide thereof is provided, wherein R1-R4 are as defined in the text. Also provided is a pharmaceutical composition comprising one or more of the described compounds of structural formula (I) and a therapeutic and/or prophylactic agent. In some embodiments, the pharmaceutical composition further comprises one or more components selected from neutral lipids, charged lipids, steroids, and polymer-conjugated lipids. Such a composition is useful for forming lipid nanoparticles for delivery of therapeutic and/or prophylactic agents. In other embodiments, the present invention provides a method of administering a therapeutic and/or prophylactic agent to a subject in need thereof, comprising preparing a pharmaceutical composition comprising lipid nanoparticles of the compound of structural formula (I) and a therapeutic and/or prophylactic agent, and delivering the composition to a subject.

Description

新型阳离子脂质化合物Novel Cationic Lipid Compounds 技术领域technical field
本发明提供新型阳离子脂质，其可用于与其他脂质组分(如中性脂质、类固醇和聚合物缀合的脂质)结合，以便形成一种核酸mRNA脂质纳米粒子组合物将一种或多种治疗剂和/或预防剂递送至哺乳动物细胞或器官和/或在哺乳动物细胞或器官中产生多肽的方法。除新型脂质外，本发明的脂质纳米粒子组合物还可以包括呈特定比例的一种或多种阳离子性和/或可离子化氨基脂质、包括多不饱和脂质在内的中性脂质、聚合物缀合的脂质、类固醇、和/或治疗剂和/或预防剂。The present invention provides novel cationic lipids that can be used in combination with other lipid components (such as neutral lipids, steroids, and polymer-conjugated lipids) to form a nucleic acid mRNA lipid nanoparticle composition combining a A method of delivering one or more therapeutic and/or prophylactic agents to a mammalian cell or organ and/or producing a polypeptide in a mammalian cell or organ. In addition to the novel lipids, the lipid nanoparticle compositions of the invention may also include, in specified proportions, one or more cationic and/or ionizable amino lipids, neutral lipids including polyunsaturated lipids, Lipids, polymer-conjugated lipids, steroids, and/or therapeutic and/or prophylactic agents.
背景技术Background technique
生物活性物质如小分子药物、蛋白质和核酸的有效靶向递送提出了一个持久的医学难题。确切地说，将核酸递送至细胞因这些物种的相对不稳定性和低细胞渗透性而变得困难。因此，需要开发有助于将治疗剂和/或预防剂如核酸递送至细胞的方法和组合物。Efficient targeted delivery of bioactive substances such as small molecule drugs, proteins and nucleic acids presents a persistent medical problem. Specifically, delivery of nucleic acids to cells is made difficult by the relative instability and low cell permeability of these species. Accordingly, there is a need to develop methods and compositions that facilitate the delivery of therapeutic and/or prophylactic agents, such as nucleic acids, to cells.
经研究证明，利用含脂质的纳米颗粒组合物、脂质体和脂质体复合物作运输媒介，可有效地将生物活性物质如小分子药物、蛋白质和核酸运送至细胞和/或细胞内隔室中。这些组合物一般包含一种或多种"阳离子性"脂质，包括多不饱和脂质在内的中性脂质(如磷脂)、结构性脂质(如类固醇)和/或含聚乙二醇的脂质(聚合物缀合的脂质)。阳离子脂质包括如可容易地质子化的含胺脂质。Studies have proved that using lipid-containing nanoparticle compositions, liposomes and liposome complexes as transport media can effectively deliver biologically active substances such as small molecule drugs, proteins and nucleic acids to cells and/or intracellularly in the compartment. These compositions generally comprise one or more "cationic" lipids, neutral lipids including polyunsaturated lipids (such as phospholipids), structured lipids (such as steroids) and/or polyethylene glycol-containing Lipids of alcohols (polymer-conjugated lipids). Cationic lipids include, for example, amine-containing lipids that can be readily protonated.
然而，在治疗环境中使用寡核苷酸目前面临着两个问题。第一，游离的RNA易于在血浆中核酸酶消化。第二，游离RNA进入存在相关翻译机制的细胞内隔室的能力受限。由阳离子脂质与其他脂质组分(如中性脂质、胆固醇、PEG、PEG化的脂质和寡核苷酸)形成的脂质纳米颗粒已用于阻止RNA在血浆中的降解并促进寡核苷酸的细胞摄取。However, the use of oligonucleotides in a therapeutic setting currently faces two problems. First, free RNA is readily digested by nucleases in plasma. Second, cell-free RNA has limited access to intracellular compartments where associated translation machinery exists. Lipid nanoparticles formed from cationic lipids with other lipid components such as neutral lipids, cholesterol, PEG, PEGylated lipids, and oligonucleotides have been used to prevent RNA degradation in plasma and promote Cellular uptake of oligonucleotides.
仍然有必要改进的用于递送寡核苷酸的阳离子脂质和脂质纳米颗粒。改进后的脂质纳米颗粒会提供优化的药物递送，保护核酸不在血清中被降解和清除，其适于全身或局部递送，并且提供核酸的细胞内递送。另外，这些优选的脂质-核酸颗粒应当是耐受良好的，并且提供足够的治疗指数，使得在有效剂量的核酸下的患者治疗不会对患者产生不可接受的毒性和/或风险。本发明提供这些优点和相关的优点。There remains a need for improved cationic lipids and lipid nanoparticles for the delivery of oligonucleotides. The improved lipid nanoparticles would provide optimized drug delivery, protect nucleic acids from degradation and clearance in serum, be suitable for systemic or local delivery, and provide intracellular delivery of nucleic acids. In addition, these preferred lipid-nucleic acid particles should be well tolerated and provide a sufficient therapeutic index such that treatment of a patient at an effective dose of the nucleic acid does not create unacceptable toxicity and/or risk to the patient. The present invention provides these and related advantages.
公开内容public content
本发明提供以下新型化合物和涉及这些化合物的方法：The present invention provides the following novel compounds and methods involving these compounds:
第一方面，本发明涉及以下结构式(I)的化合物：In a first aspect, the present invention relates to compounds of the following structural formula (I):
Figure PCTCN2022084347-appb-000001
Figure PCTCN2022084347-appb-000001
或其N-氧化物，或其盐或异构体。or its N-oxide, or its salt or isomer.
其中结构式“I”中的R 1、R 2、R 3、和R 4均为独立的2个“氢”同位素(包括同 Wherein R 1 , R 2 , R 3 , and R 4 in the structural formula "I" are two independent "hydrogen" isotopes (including isotopes
位素“氕”和“氘”)的组合，具体表现为“HH”、“HD”和“DD”的组合；combination of isotopes "protium" and "deuterium"), embodied as a combination of "HH", "HD" and "DD";
R 1和R 2独立地选自由“HH”、“HD”和“DD”组成的任意一组； R and R are independently selected from any group consisting of "HH", "HD" and "DD";
R 3和R 4独立地选自由“HH、HD”、“HD、HH”、“HH、DD”“DD、HH”“HD、DD”和“DD、 DD”组成的任意一组； R3 and R4 are independently selected from any group consisting of "HH, HD", "HD, HH", "HH, DD", "DD, HH", "HD, DD" and "DD, DD";
R 1、R 2、R 3、和R 4均有独立性，但不能同时为“HH”，即至少含有一个“HD”； R 1 , R 2 , R 3 , and R 4 are all independent, but cannot be "HH" at the same time, that is, contain at least one "HD";
在各个不同的实施方案中，所述化合物具有以下表1中所示的结构之一In various embodiments, the compound has one of the structures shown in Table 1 below
表1代表性化合物Table 1 Representative Compounds
Figure PCTCN2022084347-appb-000002
Figure PCTCN2022084347-appb-000002
Figure PCTCN2022084347-appb-000003
Figure PCTCN2022084347-appb-000003
Figure PCTCN2022084347-appb-000004
Figure PCTCN2022084347-appb-000004
Figure PCTCN2022084347-appb-000005
Figure PCTCN2022084347-appb-000005
Figure PCTCN2022084347-appb-000006
Figure PCTCN2022084347-appb-000006
Figure PCTCN2022084347-appb-000007
Figure PCTCN2022084347-appb-000007
Figure PCTCN2022084347-appb-000008
Figure PCTCN2022084347-appb-000008
Figure PCTCN2022084347-appb-000009
Figure PCTCN2022084347-appb-000009
Figure PCTCN2022084347-appb-000010
Figure PCTCN2022084347-appb-000010
Figure PCTCN2022084347-appb-000011
Figure PCTCN2022084347-appb-000011
Figure PCTCN2022084347-appb-000012
Figure PCTCN2022084347-appb-000012
在一些实施方案中，提供了包含结构式(I)的化合物中的任一种或多种和治疗剂和/或预防剂的组合物。In some embodiments, there is provided a composition comprising any one or more of the compounds of structural formula (I) and a therapeutic and/or prophylactic agent.
在一些实施方案中，提供了包含结构(I)的化合物中的任一种或多种和治疗剂和/或预防剂的组合物。在一些实施方案中，所述组合物包含结构(I)的化合物中的任一种和治疗剂和/或预防剂以及一种或多种选自中性脂质、类固醇和聚合物缀合的脂质的赋形剂。其他药物可接受的赋形剂和/或载体也包括在组合物的各种实施方案内。In some embodiments, there is provided a composition comprising any one or more of the compounds of structure (I) and a therapeutic and/or prophylactic agent. In some embodiments, the composition comprises any of the compounds of structure (I) and a therapeutic and/or prophylactic agent and one or more compounds selected from the group consisting of neutral lipids, steroids, and polymer conjugates. Lipid excipients. Other pharmaceutically acceptable excipients and/or carriers are also included in various embodiments of the compositions.
在一些实施方案中，中性脂质选自1，2-二硬脂酰基-sn-甘油-3-磷酸胆碱(DSPC)、1，2-二棕榈酰基-sn-甘油-3-磷酸胆碱(DPPC)、1，2-二肉豆寇基-sn-甘油-磷酸胆碱(DMPC)、1，2-二油酰基-sn-甘油-3-磷酸胆碱(DOPC)、1-棕榈酰基-2-油酰基-sn-甘油-3-磷酸胆碱(POPC)、1，2-二油酰基-sn-甘油-3-磷酸乙醇胺(DOPE)、鞘磷脂(SM)和其混合。在一些实施方案中，优选中性脂质为1，2-二硬脂酰基-sn-甘油-3-磷酸胆碱(DSPC)。In some embodiments, the neutral lipid is selected from 1,2-distearoyl-sn-glycero-3-phosphocholine (DSPC), 1,2-dipalmitoyl-sn-glycero-3-phosphocholine Alkaline (DPPC), 1,2-Dimyristyl-sn-glycero-phosphocholine (DMPC), 1,2-dioleoyl-sn-glycero-3-phosphocholine (DOPC), 1-palm Acyl-2-oleoyl-sn-glycero-3-phosphoethanolamine (POPC), 1,2-dioleoyl-sn-glycero-3-phosphoethanolamine (DOPE), sphingomyelin (SM) and mixtures thereof. In some embodiments, the preferred neutral lipid is 1,2-distearoyl-sn-glycero-3-phosphocholine (DSPC).
在一些实施方案中，类固醇选自胆固醇、粪固醇、谷固醇、麦角固醇、菜油固醇、豆固醇、菜籽固醇、番茄碱、熊果酸、α-生育酚和其混合。在一些实施方案中，优选类固醇为胆固醇。In some embodiments, the steroid is selected from the group consisting of cholesterol, fecal sterol, sitosterol, ergosterol, campesterol, stigmasterol, brassicasterol, tomatidine, ursolic acid, alpha-tocopherol, and mixtures thereof . In some embodiments, the preferred steroid is cholesterol.
在一些实施方案中，聚乙二醇化脂质为1,2-二肉豆蔻酰基-sn-甘油甲氧基聚乙二醇(PEG-DMG)In some embodiments, the pegylated lipid is 1,2-dimyristoyl-sn-glycerol methoxypolyethylene glycol (PEG-DMG)
在一些实施方案中，所述组合物比例以下范围：约10～60mol％所述化合物、约0～30mol％中性脂质、约10～55mol％类固醇和约0～10mol％聚合物缀合的脂质。In some embodiments, the composition ratio ranges from about 10 to 60 mol% of the compound, about 0 to 30 mol% of neutral lipids, about 10 to 55 mol% of steroids, and about 0 to 10 mol% of polymer-conjugated lipids. quality.
在一些前述组合物的实施方案中，治疗剂和/或预防剂包括核酸。其中核酸为RNA，其选自以下组成：siRNA、aiRNA、miRNA、dsRNA、shRNA、mRNA以及其混合物。在一些实施方案中，所述RNA选自mRNA。In embodiments of some of the foregoing compositions, the therapeutic and/or prophylactic agent comprises a nucleic acid. Wherein the nucleic acid is RNA, which is selected from the following composition: siRNA, aiRNA, miRNA, dsRNA, shRNA, mRNA and mixtures thereof. In some embodiments, the RNA is selected from mRNA.
在其他不同的实施方案中，本发明涉及向有需要的受试者施用治疗剂和/或预防剂的方法，该方法包括制备或提供上述组合物中的任一种并向受试者施用该组合物。In other various embodiments, the present invention relates to methods of administering therapeutic and/or prophylactic agents to a subject in need thereof, the method comprising preparing or providing any of the compositions described above and administering the composition to the subject combination.
出于施用的目的，本发明的化合物(通常是脂质纳米颗粒与治疗剂和/或预防剂结合的形式)可以以原料药施用，或者可以配制为药物组合物。本发明的药物组合物包含结构(I)的化合物和一种或多种药物可接受的载体、稀释剂或赋形剂。结构(I)的化合物以有效形成脂质纳米颗粒并递送治疗剂和/或预防剂。本领域技术人员可以容易地确定适当的浓度和剂量。For purposes of administration, the compounds of the invention, typically in the form of lipid nanoparticles in combination with therapeutic and/or prophylactic agents, can be administered as a drug substance, or can be formulated as a pharmaceutical composition. The pharmaceutical composition of the present invention comprises a compound of structure (I) and one or more pharmaceutically acceptable carriers, diluents or excipients. Compounds of structure (I) are effective to form lipid nanoparticles and deliver therapeutic and/or prophylactic agents. Appropriate concentrations and dosages can be readily determined by those skilled in the art.
本发明组合物的施用可以通过任何用于类似效用的试剂的可接受施用方式来进行。本发明的药物组合物可以配制成固体、半固体、液体或气体形式的制剂，例如片剂、胶囊、粉末、颗粒、软膏、溶液、悬浮液、栓剂、注射剂、吸入剂、凝胶、微球和气溶胶。施用这类药物组合物的典型途径包括，但不限于，口服、局部、经皮、吸入、胃肠外、舌下、口含、直肠、阴道和鼻内途径。本文使用的术语胃肠外包括皮下注射，静脉内、肌内、皮内、胸骨内注射或输注技术。配制本发明的药物组合物以便允许经过对受试者施用该组合物后其中含有的活性成分是生物可利用的。待向对象或患者施用的组合物是一个或多个剂量单位的形式，其中，片剂可以是单剂量单位，而本发明气溶胶形式的化合物的容器可以容纳多个剂量单位。制备这些剂型的现行的方法是己知的，或者对于本领域技术人员是显而易见的。在任何情况下，待施用的组合物将会含有治疗有效量的本发明化合物或其药物可接受的盐，以便根据本发明的教导治疗相关的疾病或病况。Administration of the compositions of the present invention may be by any of the acceptable modes of administration for agents of similar utility. The pharmaceutical composition of the present invention can be formulated into preparations in solid, semi-solid, liquid or gaseous form, such as tablets, capsules, powders, granules, ointments, solutions, suspensions, suppositories, injections, inhalants, gels, microspheres and aerosols. Typical routes of administration of such pharmaceutical compositions include, but are not limited to, oral, topical, transdermal, inhalation, parenteral, sublingual, buccal, rectal, vaginal and intranasal routes. The term parenteral as used herein includes subcutaneous injections, intravenous, intramuscular, intradermal, intrasternal injection or infusion techniques. The pharmaceutical compositions of the invention are formulated so as to allow the active ingredient contained therein to be bioavailable upon administration of the composition to a subject. Compositions to be administered to a subject or patient are in the form of one or more dosage units, where a tablet may be a single dosage unit and a container of the compound in aerosol form of the invention may hold multiple dosage units. Current methods for the preparation of such dosage forms are known, or will be apparent, to those skilled in the art. In any event, the composition to be administered will contain a therapeutically effective amount of a compound of the invention, or a pharmaceutically acceptable salt thereof, for treating the associated disease or condition in accordance with the teachings of the invention.
本发明的药物组合物可以是固体或液体的形式。一方面，载体是微粒，使得组合物是片剂或粉末形式。载体可以是液体，此时组合物是口服糖浆或可注射液体或气溶胶，所述气溶胶适用于吸入施用。The pharmaceutical compositions of the present invention may be in solid or liquid form. In one aspect, the carrier is particulate, such that the composition is in tablet or powder form. The carrier can be a liquid in which case the composition is an oral syrup or an injectable liquid or an aerosol suitable for administration by inhalation.
当意图用于口服施用时，药物组合物优选为固体或液体形式，其中本文认为是固体或液体的形式包 括半固体、半液体、悬浮液和凝胶形式。When intended for oral administration, the pharmaceutical composition is preferably in solid or liquid form, where forms considered solid or liquid herein include semi-solid, semi-liquid, suspension and gel forms.
作为用于口服施用的固体组合物，药物组合物可以配制成粉末、颗粒、压缩的片剂、丸剂、胶囊、咀嚼胶、薄片等形式。这类固体组合物通常将含有一种或多种惰性稀释剂或可食用载体。另外，可以存在以下的一种或多种:粘合剂，如明胶、纤维素等；赋形剂，如乳糖等；崩解剂，如海藻酸等；润滑剂，如硬脂酸镁等；助流剂，如硅胶等；甜味剂，如蔗糖或糖精；调昧剂，如薄荷等；以及着色剂。As solid compositions for oral administration, the pharmaceutical compositions can be formulated in the form of powders, granules, compressed tablets, pills, capsules, chewing gums, flakes and the like. Such solid compositions will generally contain one or more inert diluents or edible carriers. In addition, there may be one or more of the following: binders, such as gelatin, cellulose, etc.; excipients, such as lactose, etc.; disintegrants, such as alginic acid, etc.; lubricants, such as magnesium stearate, etc.; Glidants, such as silica gel, etc.; sweeteners, such as sucrose or saccharin; flavoring agents, such as mint; and coloring agents.
当药物组合物是胶囊形式时，其可以含有上述类型材料之外的液体载体，如聚乙二醇或油。When the pharmaceutical composition is in capsule form, it may contain liquid carriers other than materials of the above type, such as polyethylene glycol or oil.
药物组合物可以是液体的形式，如糖浆、溶液、乳液或悬浮液。作为两种实例，液体可以用于口服施用或用于注射递送。当意图用于口服施用时，优边的组合物含有，除本发明化合物之外的，甜味剂、防腐剂、染色/着色剂和增昧剂中的一种或多种。在通过注射施用的组合物中，可以包括表面活性剂、防腐剂、润湿剂、分散剂、悬浮剂、缓冲剂、稳定剂和等渗剂中的一种或多种。Pharmaceutical compositions may be in liquid form, such as syrups, solutions, emulsions or suspensions. Liquids can be used for oral administration or for injectable delivery, as two examples. When intended for oral administration, the compositions of Urim contain, in addition to the compounds of the present invention, one or more of sweetening agents, preservatives, dyeing/coloring agents and darkening agents. In compositions administered by injection, one or more of surfactants, preservatives, wetting agents, dispersing agents, suspending agents, buffers, stabilizers and isotonic agents may be included.
本发明的液体药物组合物，不论其为溶液、悬浮液还是其他类似的形式，可以包括以下佐剂中的一种或多种:无菌稀释剂，如注射用水、盐水溶液、优选生理盐水、林格氏溶液、等渗氯化钠；不挥发性油类，如可用作溶剂或悬浮介质的合成的单甘酯或双甘酯，聚乙二醇、甘油、丙二醇或其他溶剂；抗菌剂，如尼泊金甲醋等；抗氧化剂，如抗坏血酸或亚硫酸氢钠；螯合剂，如乙二肢四乙酸；缓冲剂，如乙酸盐、拧棱酸盐或磷酸盐；以及用于调节张力的试剂，如氯化钠或葡萄糖；用作冷冻保护剂的试剂，如蔗糖或海藻糖。胃肠外制剂可以封装在玻璃或塑料制作的安瓿、一次性注射器或多剂量瓶中。生理盐水是优选的佐剂。可注射的药物组合物优选为无菌的。The liquid pharmaceutical composition of the present invention, whether it is a solution, a suspension or other similar forms, may include one or more of the following adjuvants: sterile diluents, such as water for injection, saline solution, preferably physiological saline, Ringer's solution, isotonic sodium chloride; fixed oils such as synthetic mono- or diglycerides, polyethylene glycol, glycerol, propylene glycol, or other solvents that may be used as a solvent or suspending medium; antibacterial agents , such as methylparaben, etc.; antioxidants, such as ascorbic acid or sodium bisulfite; chelating agents, such as ethylenedipic acid; buffering agents, such as acetate, clematine, or phosphate; Reagents for tonicity, such as sodium chloride or glucose; reagents used as cryoprotectants, such as sucrose or trehalose. The parenteral preparation can be enclosed in ampoules, disposable syringes or multiple dose vials made of glass or plastic. Physiological saline is the preferred adjuvant. Injectable pharmaceutical compositions are preferably sterile.
本发明的药物组合物可以意图用于局部施用，在这种情况下，载体可以适当地包含溶液基质、乳液基质、软膏基质或疑胶基质。所述基质可以包含以下的一种或多种：矿脂、羊毛脂、聚乙二醇、蜂蜡、矿物油、诸如水和醇的稀释剂、以及乳化剂和稳定剂。增稠剂可以存在于用于局部施用的药物组合物中。如果意图用于经皮施用，组合物可以包括经皮贴片或离子电渗装置。The pharmaceutical compositions of the present invention may be intended for topical administration, in which case the carrier may suitably comprise a solution base, emulsion base, ointment base or gel base. The base may contain one or more of petrolatum, lanolin, polyethylene glycols, beeswax, mineral oil, diluents such as water and alcohol, and emulsifiers and stabilizers. Thickening agents may be present in pharmaceutical compositions for topical administration. If intended for transdermal administration, the composition may include a transdermal patch or iontophoretic device.
本发明的药物组合物可以包括修饰固体或液体剂量单位的物理形式的各种材料。组合物可以包括形成活性成分周围的包衣壳的材料。形成包衣壳的材料通常是惰性的，并且可以为，糖、虫胶、和其他肠溶包衣试剂。或者，活性成分可以封装在明胶胶囊中。The pharmaceutical compositions of the invention may include various materials which modify the physical form of solid or liquid dosage units. The composition may include materials which form a coating shell around the active ingredient. Materials forming the coating shell are generally inert and can be, sugar, shellac, and other enteric coating agents. Alternatively, the active ingredient may be enclosed in a gelatin capsule.
固体或液体形式的本发明药物组合物可以包括与本发明化合物结合并由此有助于所述化合物递送的试剂。可以以这种能力作用的适合的试剂包括单克隆或多克隆抗体或蛋白质。Pharmaceutical compositions of the invention in solid or liquid form may include agents that bind to and thereby facilitate delivery of the compounds of the invention. Suitable agents that may act in this capacity include monoclonal or polyclonal antibodies or proteins.
本发明的药物组合物可以由可作为气溶胶施用的剂量单位组成。术语气溶胶用于表示从胶体性质的系统到由加压包装组成的系统的各种系统。可以通过液化气或压缩气来递送，或者通过分散活性成分的适合的泵系统来递送。本发明化合物的气溶胶可以以单相、双相系统或三相系统来递送，以便递送活性成分。气溶胶的递送包括必要的容器、活化剂、阀、子容器等，其在一起可以形成试剂盒。本领域技术人员不需过度实验即可确定优选的气溶胶。The pharmaceutical compositions of the invention may consist of dosage units which can be administered as an aerosol. The term aerosol is used to denote systems ranging from those of a colloidal nature to those consisting of pressurized packs. Delivery may be by liquefied or compressed gas, or by a suitable pump system which dispenses the active ingredient. Aerosols of the compounds of the invention may be delivered as single-phase, bi-phasic or triphasic systems to deliver the active ingredient. Aerosol delivery includes the necessary containers, activators, valves, sub-containers, etc., which together may form a kit. Preferred aerosols can be determined by those skilled in the art without undue experimentation.
本发明的药物组合物可以通过制药领域熟知的方法来制备。意图通过注射施用的药物组合物可以通过将本发明的脂质纳米颗粒与无菌的蒸馏水或其他载体结合成溶液来制备。可以加入表面活性剂以促进形成均匀的溶液或悬浮液。表面活性剂是与本发明化合物非共价地相互作用，以便促进所述化合物在水性递送系统中溶解或均匀悬浮的化合物。The pharmaceutical compositions of the present invention can be prepared by methods well known in the field of pharmacy. Pharmaceutical compositions intended to be administered by injection can be prepared by combining the lipid nanoparticles of the present invention with sterile distilled water or other carriers into a solution. Surfactants can be added to promote the formation of a uniform solution or suspension. Surfactants are compounds that interact non-covalently with the compounds of the invention in order to facilitate dissolution or uniform suspension of the compounds in aqueous delivery systems.
本发明的组合物或其药物可接受的盐以治疗有效量施用，所述量将会根据多种因素变化，包括使用的具体治疗剂的活性；治疗剂的代谢稳定性和作用时长；受试者的年龄、体重、一般健康状况、性别和饮食；施用的方式和时间；排泄速率；药物组合；具体病例的严重性等。The composition of the present invention, or a pharmaceutically acceptable salt thereof, is administered in a therapeutically effective amount, which will vary depending on a number of factors, including the activity of the particular therapeutic agent used; the metabolic stability and duration of action of the therapeutic agent; The patient's age, weight, general health, sex, and diet; mode and timing of administration; rate of excretion; drug combination; severity of the specific case, etc.
本发明的组合物也可以在施用一种或多种其他治疗剂的同时、之前或之后施用。这类组合治疗包括施用本发明组合物和一种或多种另外的活性剂的单一药物剂量制剂，以及施用本发明组合物和各个在其自身单独药物剂量制剂中的活性剂。例如，本发明组合物和其他活性剂可以以单一口服剂量组合物(如片剂或胶囊)一起向受试者施用，或者各个试剂以不同的口服剂量制剂施用。当使用不同的剂量制剂时，本发明化合物和一种或多种另外的活性剂可以在基本同一时间施用，或者在相互交错的时间依次施用；应理解组合治疗包括所有的这些给药方案。Compositions of the invention may also be administered concurrently with, before or after administration of one or more other therapeutic agents. Such combination therapy includes administration of a single pharmaceutical dosage formulation of a composition of the invention and one or more additional active agents, as well as administration of a composition of the invention and each active agent in its own separate pharmaceutical dosage formulation. For example, compositions of the invention and other active agents may be administered to a subject together in a single oral dosage composition such as a tablet or capsule, or each agent may be administered in separate oral dosage formulations. When different dosage formulations are used, the compound of the invention and the one or more additional active agents may be administered at substantially the same time, or sequentially at times staggered from each other; it is understood that combination therapy includes all such dosing regimens.
上述新型氘代阳离子脂质化合物的结构修饰和设计，实现了更有优势的理化性质，包括更合适的pKa和更好的化学稳定性，用于mRNA纳米脂质体组合物，可实现对离子类核酸药物更有效结合并递送，同时其化学结构更稳定，便于合成和有利开发为药用辅料。The structural modification and design of the above-mentioned novel deuterated cationic lipid compounds have achieved more advantageous physical and chemical properties, including more suitable pKa and better chemical stability, and are used in mRNA nanoliposome compositions, which can realize anti-ion Nucleic acid-like drugs are more effectively combined and delivered, and at the same time, their chemical structure is more stable, which is convenient for synthesis and beneficial for development as pharmaceutical excipients.
上述化合物和组合物的制备方法在下文描述，和/或在本领域已知。Methods for the preparation of the above compounds and compositions are described below, and/or are known in the art.
本领域的技术人员将会认识到，在本文描述的方法中，中间化合物的官能团可能需要通过适合的保护基保护。这类官能团包括羟基、氨基和羧酸。用于羟基的适合的保护基包括三烷基甲硅烷基或二芳基烷基甲硅烷基、四氢呋喃基、苄基等。用于氨基的适合的保护基包括叔丁氧基羰基、苄氧基羰基等。用于羧酸的适合的保护基包括烃基、芳基或芳烃基酯。保护基可以根据标准技术添加或去除，所述标准技术是本领域技术人员已知的合本文中描述的。Those skilled in the art will recognize that in the methods described herein, functional groups of intermediate compounds may need to be protected by suitable protecting groups. Such functional groups include hydroxyl, amino and carboxylic acid. Suitable protecting groups for hydroxy include trialkylsilyl or diarylalkylsilyl, tetrahydrofuranyl, benzyl, and the like. Suitable protecting groups for amino include t-butoxycarbonyl, benzyloxycarbonyl and the like. Suitable protecting groups for carboxylic acids include hydrocarbyl, aryl or arylhydrocarbyl esters. Protecting groups can be added or removed according to standard techniques, known to those skilled in the art and described herein.
本领域技术人员还将认识到，虽然本发明化合物的这类经保护的衍生物可以不由此具有药物活性，但其可以对哺乳动物施用并且之后在体内代谢形成具有药理活性的本发明化合物。这类衍生物因此可以被描述为“前药”。所以本发明化合物的前药包括在本发明的范围内。Those skilled in the art will also recognize that while such protected derivatives of compounds of the invention may not thereby be pharmaceutically active, they may be administered to a mammal and then metabolized in vivo to form pharmacologically active compounds of the invention. Such derivatives can thus be described as "prodrugs". Prodrugs of the compounds of the present invention are therefore included within the scope of the present invention.
此外，所有以游离碱或游离酸形式存在的本发明化合物可以根据本领域技术人员已知的方法用适当的无机或有机的碱或酸处理来转化为其药物可接受的盐。本发明化合物的盐可以通过标准技术转化为其游离碱或酸形成。In addition, all compounds of the present invention that exist in free base or free acid form can be converted to their pharmaceutically acceptable salts by treatment with an appropriate inorganic or organic base or acid according to methods known to those skilled in the art. Salts of compounds of the present invention may be converted to their free base or acid forms by standard techniques.
提供了以下实施例，其目的在于举例展示，并非限定。The following examples are offered for purposes of illustration and not limitation.
以下实施例，除非另外指出，否则使用的所有溶剂和试剂都是商购得到并且以原样使用。In the following examples, unless otherwise indicated, all solvents and reagents used were obtained commercially and used as received.
以下描述的程序可用于合成表1中的化合物I。The procedure described below was used to synthesize Compound I in Table 1.
本文采用了以下缩写:This article uses the following abbreviations:
DCM：二氯甲烷DCM: dichloromethane
EtOH:乙醇EtOH: ethanol
K 2CO 3:碳酸钠 K 2 CO 3 : Sodium Carbonate
EA:乙酸乙酯EA: ethyl acetate
DMSO:二甲基亚砜DMSO: dimethyl sulfoxide
具体实施方式Detailed ways
实施例1：Example 1:
代表性路线representative route
化合物8D的合成Synthesis of compound 8D
Figure PCTCN2022084347-appb-000013
Figure PCTCN2022084347-appb-000013
1)化合物C的合成1) Synthesis of Compound C
Figure PCTCN2022084347-appb-000014
Figure PCTCN2022084347-appb-000014
25ml双口瓶中加入NH3(1g,.7M in MeOH)和THF,加入溴乙醇(4eq.),温到65℃反应6小时，过滤，得到类白色固体1.5g目标化合物，收率75％Add NH3 (1g,.7M in MeOH) and THF to a 25ml two-necked bottle, add bromoethanol (4eq.), warm to 65°C and react for 6 hours, filter to obtain 1.5g of the target compound as an off-white solid, yield 75%
化学式：C6H15NO3Chemical formula: C6H15NO3
分子量：149.19Molecular weight: 149.19
LC-MS:m/z 150[M+H]LC-MS: m/z 150[M+H]
H NMR(300MHz,DMSO-d6):1H NMR(DMSO-d6):δ(ppm)7.31(s,1H)4.16(3H,s),3.99(t,6H,J＝6)3.48(t,6H,J＝8).H NMR(300MHz,DMSO-d6):1H NMR(DMSO-d6):δ(ppm)7.31(s,1H)4.16(3H,s),3.99(t,6H,J＝6)3.48(t,6H ,J＝8).
2)化合物8D-4的合成2) Synthesis of Compound 8D-4
Figure PCTCN2022084347-appb-000015
Figure PCTCN2022084347-appb-000015
25ml的双口瓶中加入1.5g 8D-3(10.05mmol),加入15ml氯仿，加入SOCl2(13.07mmol,1.6g),室温搅拌3小时，用碳酸氢钠调pH＝8-9，用30ml DCM萃取1次，用6ml食盐水洗一次，有机相浓缩得到黄色液体，加入10ml THF，加入1.3当量盐酸，成盐析晶，室温搅拌5小时，过滤得到1.7g白色固体，收率70％。Add 1.5g 8D-3 (10.05mmol) to a 25ml two-necked bottle, add 15ml chloroform, add SOCl2 (13.07mmol, 1.6g), stir at room temperature for 3 hours, adjust the pH to 8-9 with sodium bicarbonate, and use 30ml DCM Extract once, wash once with 6ml of salt water, concentrate the organic phase to obtain a yellow liquid, add 10ml of THF, add 1.3 N hydrochloric acid, form salt and crystallize, stir at room temperature for 5 hours, filter to obtain 1.7g of white solid, yield 70%.
化学式：C 6H 12NCl 3.HCl Chemical formula: C 6 H 12 NCl 3 .HCl
分子量：241.02Molecular weight: 241.02
LC-MS:m/z 205[M+H]LC-MS: m/z 205[M+H]
H NMR(300MHz,DMSO-d6):1H NMR(DMSO-d6):δ(ppm)7.25(s,1H),3.82(t,6H,J＝4)3.54(t,6H,J＝7).H NMR (300MHz, DMSO-d6): 1H NMR (DMSO-d6): δ (ppm) 7.25 (s, 1H), 3.82 (t, 6H, J＝4) 3.54 (t, 6H, J＝7).
3)化合物8D-6的合成3) Synthesis of Compound 8D-6
Figure PCTCN2022084347-appb-000016
Figure PCTCN2022084347-appb-000016
25ml的双口瓶中加入1.7g 8D-4(6.45mmol),加入20ml甲苯，加入8D-5(14.84mmol,0.95g),加入KOH(45.16mmol,2.9g)，升温到115℃搅拌8小时，反应完后，降到室温，用25*3ml EA萃取3次，合并有机相用10ml食盐水洗一次，有机相浓缩得到黄色液体，加入20ml THF，加入3.5当量盐酸，成盐析晶，室温搅拌8小时，过滤得到1.29g白色固体，收率60％Add 1.7g 8D-4 (6.45mmol) to a 25ml two-necked flask, add 20ml toluene, add 8D-5 (14.84mmol, 0.95g), add KOH (45.16mmol, 2.9g), heat up to 115°C and stir for 8 hours After the reaction, cool down to room temperature, extract 3 times with 25*3ml EA, combine the organic phase and wash once with 10ml salt water, concentrate the organic phase to obtain a yellow liquid, add 20ml THF, add 3.5 equivalent hydrochloric acid, form salt and crystallize, stir at room temperature 8 hours, filtered to obtain 1.29g white solid, yield 60%
化学式：C 10H 17D 8N 5 Chemical formula: C 10 H 17 D 8 N 5
分子量：223.39Molecular weight: 223.39
LC-MS:m/z 224[M+H]LC-MS: m/z 224[M+H]
H NMR(300MHz,DMSO-d6):1H NMR(DMSO-d6):δ(ppm)8.44(s,6H),3.32(s,2H,)2.53(m,2H),2.39(m,2H)2.22(m,6H).H NMR(300MHz,DMSO-d6):1H NMR(DMSO-d6):δ(ppm)8.44(s,6H),3.32(s,2H,)2.53(m,2H),2.39(m,2H)2.22 (m,6H).
4)化合物8D的合成4) Synthesis of Compound 8D
Figure PCTCN2022084347-appb-000017
Figure PCTCN2022084347-appb-000017
25ml的双口瓶中加入0.9g 8D-6base(4.03mmol),加入8D-6(22.16mmol,4.08g)升温到80-90℃搅拌5天，反应完后，降到室温，通过硅胶色谱法(梯度洗脱为DCM到175:22:3DCM/MeOH/NH4OH(水溶液))分离所需产品，浓缩得到黄色油状物1.8g，收率40％Add 0.9g 8D-6base (4.03mmol) into a 25ml two-necked bottle, add 8D-6 (22.16mmol, 4.08g) and heat up to 80-90°C and stir for 5 days. After the reaction, cool down to room temperature and pass through silica gel chromatography. (Gradient elution from DCM to 175:22:3 DCM/MeOH/NH4OH (aq)) isolated the desired product and concentrated to give 1.8 g of yellow oil, yield 40%
化学式：C 70H 137D 8N 5O 5 Chemical formula: C 70 H 137 D 8 N 5 O 5
分子量：1145.01Molecular weight: 1145.01
LC-MS:m/z 1146[M+H]LC-MS: m/z 1146[M+H]
H NMR(300MHz,DMSO-d6):5.4(s,5H),3.49(m,5H)2.22-2.71(m,20H),1.47(m,10H),1.28(m,80H),0.90(m,10H).H NMR(300MHz,DMSO-d6):5.4(s,5H),3.49(m,5H)2.22-2.71(m,20H),1.47(m,10H),1.28(m,80H),0.90(m, 10H).
实施例2：Example 2:
化合物1的合成
Figure PCTCN2022084347-appb-000018
Synthesis of compound 1
Figure PCTCN2022084347-appb-000018
化学式：C 70H 144DN 5O 5 Chemical formula: C 70 H 144 DN 5 O 5
分子量：1138.01Molecular weight: 1138.01
化合物1可根据实施例1描述的代表性路线合成。Compound 1 can be synthesized according to the representative route described in Example 1.
H NMR(300MHz,DMSO-d6):1H NMR(DMSO-d6):δ(ppm)5.4(s,5H),3.49(m,5H,)2.22-2.71(m,27H),1.47(m,10H),1.28(m,80H),0.90(m,10H).H NMR(300MHz,DMSO-d6):1H NMR(DMSO-d6):δ(ppm)5.4(s,5H),3.49(m,5H,)2.22-2.71(m,27H),1.47(m,10H ),1.28(m,80H),0.90(m,10H).
实施例3：Example 3:
化合物2的合成Synthesis of compound 2
Figure PCTCN2022084347-appb-000019
Figure PCTCN2022084347-appb-000019
化学式：C 70H 144DN 5O 5 Chemical formula: C 70 H 144 DN 5 O 5
分子量：1138.01Molecular weight: 1138.01
化合物2可根据实施例1描述的代表性路线合成。Compound 2 can be synthesized according to the representative route described in Example 1.
H NMR(300MHz,DMSO-d6):1H NMR(DMSO-d6):δ(ppm)5.4(s,5H),3.49(m,5H,)2.22-2.71(m,27H),1.47(m,10H),1.28(m,80H),0.90(m,10H).H NMR(300MHz,DMSO-d6):1H NMR(DMSO-d6):δ(ppm)5.4(s,5H),3.49(m,5H,)2.22-2.71(m,27H),1.47(m,10H ),1.28(m,80H),0.90(m,10H).
实施例4：Example 4:
化合物3的合成Synthesis of compound 3
Figure PCTCN2022084347-appb-000020
Figure PCTCN2022084347-appb-000020
化学式：C 70H 144DN 5O 5 Chemical formula: C 70 H 144 DN 5 O 5
分子量：1138.01Molecular weight: 1138.01
化合物3可根据实施例1描述的代表性路线合成。Compound 3 can be synthesized according to the representative route described in Example 1.
H NMR(300MHz,DMSO-d6):1H NMR(DMSO-d6):δ(ppm)5.4(s,5H),3.49(m,5H,)2.22-2.71(m,27H),1.47(m,10H),1.28(m,80H),0.90(m,10H).H NMR(300MHz,DMSO-d6):1H NMR(DMSO-d6):δ(ppm)5.4(s,5H),3.49(m,5H,)2.22-2.71(m,27H),1.47(m,10H ),1.28(m,80H),0.90(m,10H).
实施例5：Example 5:
化合物4的合成Synthesis of Compound 4
Figure PCTCN2022084347-appb-000021
Figure PCTCN2022084347-appb-000021
化学式：C 70H 144DN 5O 5 Chemical formula: C 70 H 144 DN 5 O 5
分子量：1138.01Molecular weight: 1138.01
化合物4可根据实施例1描述的代表性路线合成。Compound 4 can be synthesized according to the representative route described in Example 1.
H NMR(300MHz,DMSO-d6):1H NMR(DMSO-d6):δ(ppm)5.4(s,5H),3.49(m,5H,)2.22-2.71(m,27H),1.47(m,10H)1.28(m,80H),0.90(m,10H).H NMR(300MHz,DMSO-d6):1H NMR(DMSO-d6):δ(ppm)5.4(s,5H),3.49(m,5H,)2.22-2.71(m,27H),1.47(m,10H )1.28(m,80H),0.90(m,10H).
实施例6：Embodiment 6:
化合物5的合成Synthesis of compound 5
Figure PCTCN2022084347-appb-000022
Figure PCTCN2022084347-appb-000022
化学式：C 70H 143D 2N 5O 5 Chemical formula: C 70 H 143 D 2 N 5 O 5
分子量：1139.01Molecular weight: 1139.01
化合物5可根据实施例1描述的代表性路线合成。Compound 5 can be synthesized according to the representative route described in Example 1.
H NMR(300MHz,DMSO-d6):1H NMR(DMSO-d6):δ(ppm)5.4(s,5H),3.49(m,5H,)2.22-2.71(m,26H),1.47(m,10H)1.28(m,80H),0.90(m,10H).H NMR(300MHz,DMSO-d6):1H NMR(DMSO-d6):δ(ppm)5.4(s,5H),3.49(m,5H,)2.22-2.71(m,26H),1.47(m,10H )1.28(m,80H),0.90(m,10H).
实施例7：Embodiment 7:
化合物6的合成Synthesis of compound 6
Figure PCTCN2022084347-appb-000023
Figure PCTCN2022084347-appb-000023
化学式：C 70H 143D 2N 5O 5 Chemical formula: C 70 H 143 D 2 N 5 O 5
分子量：1139.01Molecular weight: 1139.01
化合物6可根据实施例1描述的代表性路线合成。Compound 6 can be synthesized according to the representative route described in Example 1.
H NMR(300MHz,DMSO-d6):1H NMR(DMSO-d6):δ(ppm)5.4(s,5H),3.49(m,5H,)2.22-2.71(m,26H),1.47(m,10H)1.28(m,80H),0.90(m,10H).H NMR(300MHz,DMSO-d6):1H NMR(DMSO-d6):δ(ppm)5.4(s,5H),3.49(m,5H,)2.22-2.71(m,26H),1.47(m,10H )1.28(m,80H),0.90(m,10H).
实施例8：Embodiment 8:
化合物7的合成Synthesis of compound 7
Figure PCTCN2022084347-appb-000024
Figure PCTCN2022084347-appb-000024
化学式：C 70H 143D 2N 5O 5 Chemical formula: C 70 H 143 D 2 N 5 O 5
分子量：1139.01Molecular weight: 1139.01
化合物7可根据实施例1描述的代表性路线合成。Compound 7 can be synthesized according to the representative route described in Example 1.
H NMR(300MHz,DMSO-d6):1H NMR(DMSO-d6):δ(ppm)5.4(s,5H),3.49(m,5H,)2.22-2.71(m,26H),1.47(m,10H)1.28(m,80H),0.90(m,10H).H NMR(300MHz,DMSO-d6):1H NMR(DMSO-d6):δ(ppm)5.4(s,5H),3.49(m,5H,)2.22-2.71(m,26H),1.47(m,10H )1.28(m,80H),0.90(m,10H).
实施例9：Embodiment 9:
化合物8的合成Synthesis of Compound 8
Figure PCTCN2022084347-appb-000025
Figure PCTCN2022084347-appb-000025
化学式：C 70H 143D 2N 5O 5 Chemical formula: C 70 H 143 D 2 N 5 O 5
分子量：1139.01Molecular weight: 1139.01
化合物8可根据实施例1描述的代表性路线合成。Compound 8 can be synthesized according to the representative route described in Example 1.
H NMR(300MHz,DMSO-d6):1H NMR(DMSO-d6):δ(ppm)5.4(s,5H),3.49(m,5H,)2.22-2.71(m,26H),1.47(m,10H)1.28(m,80H),0.90(m,10H).H NMR(300MHz,DMSO-d6):1H NMR(DMSO-d6):δ(ppm)5.4(s,5H),3.49(m,5H,)2.22-2.71(m,26H),1.47(m,10H )1.28(m,80H),0.90(m,10H).
实施例10：Example 10:
化合物9的合成Synthesis of Compound 9
Figure PCTCN2022084347-appb-000026
Figure PCTCN2022084347-appb-000026
化学式：C 70H 143D 2N 5O 5 Chemical formula: C 70 H 143 D 2 N 5 O 5
分子量：1139.01Molecular weight: 1139.01
化合物9可根据实施例1描述的代表性路线合成。Compound 9 can be synthesized according to the representative route described in Example 1.
H NMR(300MHz,DMSO-d6):1H NMR(DMSO-d6):δ(ppm)5.4(s,5H),3.49(m,5H,)2.22-2.71(m,26H),1.47(m,10H)1.28(m,80H),0.90(m,10H).H NMR(300MHz,DMSO-d6):1H NMR(DMSO-d6):δ(ppm)5.4(s,5H),3.49(m,5H,)2.22-2.71(m,26H),1.47(m,10H )1.28(m,80H),0.90(m,10H).
实施例11：Example 11:
化合物10的合成Synthesis of Compound 10
Figure PCTCN2022084347-appb-000027
Figure PCTCN2022084347-appb-000027
化学式：C 70H 143D 2N 5O 5 Chemical formula: C 70 H 143 D 2 N 5 O 5
分子量：1139.01Molecular weight: 1139.01
化合物10可根据实施例1描述的代表性路线合成。Compound 10 can be synthesized according to the representative route described in Example 1.
H NMR(300MHz,DMSO-d6):1H NMR(DMSO-d6):δ(ppm)5.4(s,5H),3.49(m,5H,)2.22-2.71(m,26H),1.47(m,10H)1.28(m,80H),0.90(m,10H).H NMR(300MHz,DMSO-d6):1H NMR(DMSO-d6):δ(ppm)5.4(s,5H),3.49(m,5H,)2.22-2.71(m,26H),1.47(m,10H )1.28(m,80H),0.90(m,10H).
实施例12：Example 12:
化合物11的合成Synthesis of Compound 11
Figure PCTCN2022084347-appb-000028
Figure PCTCN2022084347-appb-000028
化学式：C 70H 143D 2N 5O 5 Chemical formula: C 70 H 143 D 2 N 5 O 5
分子量：1139.01Molecular weight: 1139.01
化合物11可根据实施例1描述的代表性路线合成。Compound 11 can be synthesized according to the representative route described in Example 1.
H NMR(300MHz,DMSO-d6):1H NMR(DMSO-d6):δ(ppm)5.4(s,5H),3.49(m,5H,)2.22-2.71(m,26H),1.47(m,10H)1.28(m,80H),0.90(m,10H).H NMR(300MHz,DMSO-d6):1H NMR(DMSO-d6):δ(ppm)5.4(s,5H),3.49(m,5H,)2.22-2.71(m,26H),1.47(m,10H )1.28(m,80H),0.90(m,10H).
实施例13：Example 13:
化合物12的合成Synthesis of Compound 12
Figure PCTCN2022084347-appb-000029
Figure PCTCN2022084347-appb-000029
化学式：C 70H 143D 2N 5O 5 Chemical formula: C 70 H 143 D 2 N 5 O 5
分子量：1139.01Molecular weight: 1139.01
化合物12可根据实施例1描述的代表性路线合成。Compound 12 can be synthesized according to the representative route described in Example 1.
H NMR(300MHz,DMSO-d6):1H NMR(DMSO-d6):δ(ppm)5.4(s,5H),3.49(m,5H,)2.22-2.71(m,26H),1.47(m,10H)1.28(m,80H),0.90(m,10H).H NMR(300MHz,DMSO-d6):1H NMR(DMSO-d6):δ(ppm)5.4(s,5H),3.49(m,5H,)2.22-2.71(m,26H),1.47(m,10H )1.28(m,80H),0.90(m,10H).
实施例14：Example 14:
化合物13的合成Synthesis of Compound 13
Figure PCTCN2022084347-appb-000030
Figure PCTCN2022084347-appb-000030
化学式：C 70H 143D 2N 5O 5 Chemical formula: C 70 H 143 D 2 N 5 O 5
分子量：1139.01Molecular weight: 1139.01
化合物13可根据实施例1描述的代表性路线合成。Compound 13 can be synthesized according to the representative route described in Example 1.
H NMR(300MHz,DMSO-d6):1H NMR(DMSO-d6):δ(ppm)5.4(s,5H),3.49(m,5H,),2.22-2.71(m,26H),1.47(m,10H),1.28(m,80H),0.90(m,10H).H NMR(300MHz,DMSO-d6):1H NMR(DMSO-d6):δ(ppm)5.4(s,5H),3.49(m,5H,),2.22-2.71(m,26H),1.47(m, 10H),1.28(m,80H),0.90(m,10H).
实施例15：Example 15:
化合物14的合成Synthesis of Compound 14
Figure PCTCN2022084347-appb-000031
Figure PCTCN2022084347-appb-000031
化学式：C 70H 143D 2N 5O 5 Chemical formula: C 70 H 143 D 2 N 5 O 5
分子量：1139.01Molecular weight: 1139.01
化合物14可根据实施例1描述的代表性路线合成。Compound 14 can be synthesized according to the representative route described in Example 1.
H NMR(300MHz,DMSO-d6):1H NMR(DMSO-d6):δ(ppm)5.4(s,5H),3.49(m,5H,),2.22-2.71(m,26H),1.47(m,10H),1.28(m,80H),0.90(m,10H).H NMR(300MHz,DMSO-d6):1H NMR(DMSO-d6):δ(ppm)5.4(s,5H),3.49(m,5H,),2.22-2.71(m,26H),1.47(m, 10H),1.28(m,80H),0.90(m,10H).
实施例16：Example 16:
化合物15的合成Synthesis of Compound 15
Figure PCTCN2022084347-appb-000032
Figure PCTCN2022084347-appb-000032
化学式：C 70H 143D 2N 5O 5 Chemical formula: C 70 H 143 D 2 N 5 O 5
分子量：1139.01Molecular weight: 1139.01
化合物15可根据实施例1描述的代表性路线合成。Compound 15 can be synthesized according to the representative route described in Example 1.
H NMR(300MHz,DMSO-d6):1H NMR(DMSO-d6):δ(ppm)5.4(s,5H),3.49(m,5H,),2.22-2.71(m,26H),1.47(m,10H),1.28(m,80H),0.90(m,10H).H NMR(300MHz,DMSO-d6):1H NMR(DMSO-d6):δ(ppm)5.4(s,5H),3.49(m,5H,),2.22-2.71(m,26H),1.47(m, 10H),1.28(m,80H),0.90(m,10H).
实施例17：Example 17:
化合物16的合成Synthesis of compound 16
Figure PCTCN2022084347-appb-000033
Figure PCTCN2022084347-appb-000033
化学式：C 70H 142D 3N 5O 5 Chemical formula: C 70 H 142 D 3 N 5 O 5
分子量：1140.01Molecular weight: 1140.01
化合物16可根据实施例1描述的代表性路线合成。Compound 16 can be synthesized according to the representative route described in Example 1.
H NMR(300MHz,DMSO-d6):1H NMR(DMSO-d6):δ(ppm)5.4(s,5H),3.49(m,5H,),2.22-2.71(m,25H),1.47(m,10H),1.28(m,80H),0.90(m,10H).H NMR(300MHz,DMSO-d6):1H NMR(DMSO-d6):δ(ppm)5.4(s,5H),3.49(m,5H,),2.22-2.71(m,25H),1.47(m, 10H),1.28(m,80H),0.90(m,10H).
实施例18：Example 18:
化合物17的合成Synthesis of Compound 17
Figure PCTCN2022084347-appb-000034
Figure PCTCN2022084347-appb-000034
化学式：C 70H 142D 3N 5O 5 Chemical formula: C 70 H 142 D 3 N 5 O 5
分子量：1140.01Molecular weight: 1140.01
化合物17可根据实施例1描述的代表性路线合成。Compound 17 can be synthesized according to the representative route described in Example 1.
H NMR(300MHz,DMSO-d6):1H NMR(DMSO-d6):δ(ppm)5.4(s,5H),3.49(m,5H,),2.22-2.71(m,25H),1.47(m,10H),1.28(m,80H),0.90(m,10H).H NMR(300MHz,DMSO-d6):1H NMR(DMSO-d6):δ(ppm)5.4(s,5H),3.49(m,5H,),2.22-2.71(m,25H),1.47(m, 10H),1.28(m,80H),0.90(m,10H).
实施例19：Example 19:
化合物18的合成Synthesis of Compound 18
Figure PCTCN2022084347-appb-000035
Figure PCTCN2022084347-appb-000035
化学式：C 70H 142D 3N 5O 5 Chemical formula: C 70 H 142 D 3 N 5 O 5
分子量：1140.01Molecular weight: 1140.01
化合物18可根据实施例1描述的代表性路线合成。Compound 18 can be synthesized according to the representative route described in Example 1.
H NMR(300MHz,DMSO-d6):1H NMR(DMSO-d6):δ(ppm)5.4(s,5H),3.49(m,5H,),2.22-2.71(m,25H),1.47(m,10H),1.28(m,80H),0.90(m,10H).H NMR(300MHz,DMSO-d6):1H NMR(DMSO-d6):δ(ppm)5.4(s,5H),3.49(m,5H,),2.22-2.71(m,25H),1.47(m, 10H),1.28(m,80H),0.90(m,10H).
实施例20：Example 20:
化合物19的合成Synthesis of Compound 19
Figure PCTCN2022084347-appb-000036
Figure PCTCN2022084347-appb-000036
化学式：C 70H 142D 3N 5O 5 Chemical formula: C 70 H 142 D 3 N 5 O 5
分子量：1140.01Molecular weight: 1140.01
化合物19可根据实施例1描述的代表性路线合成。Compound 19 can be synthesized according to the representative route described in Example 1.
H NMR(300MHz,DMSO-d6):1H NMR(DMSO-d6):δ(ppm)5.4(s,5H),3.49(m,5H,),2.22-2.71(m,25H),1.47(m,10H),1.28(m,80H),0.90(m,10H).H NMR(300MHz,DMSO-d6):1H NMR(DMSO-d6):δ(ppm)5.4(s,5H),3.49(m,5H,),2.22-2.71(m,25H),1.47(m, 10H),1.28(m,80H),0.90(m,10H).
实施例21：Example 21:
化合物20的合成Synthesis of compound 20
Figure PCTCN2022084347-appb-000037
Figure PCTCN2022084347-appb-000037
化学式：C 70H 142D 3N 5O 5 Chemical formula: C 70 H 142 D 3 N 5 O 5
分子量：1140.01Molecular weight: 1140.01
化合物20可根据实施例1描述的代表性路线合成。Compound 20 can be synthesized according to the representative route described in Example 1.
H NMR(300MHz,DMSO-d6):1H NMR(DMSO-d6):δ(ppm)5.4(s,5H),3.49(m,5H,),2.22-2.71(m,25H),1.47(m,10H),1.28(m,80H),0.90(m,10H).H NMR(300MHz,DMSO-d6):1H NMR(DMSO-d6):δ(ppm)5.4(s,5H),3.49(m,5H,),2.22-2.71(m,25H),1.47(m, 10H),1.28(m,80H),0.90(m,10H).
实施例22：Example 22:
化合物21的合成Synthesis of Compound 21
Figure PCTCN2022084347-appb-000038
Figure PCTCN2022084347-appb-000038
化学式：C 70H 142D 3N 5O 5 Chemical formula: C 70 H 142 D 3 N 5 O 5
分子量：1140.01Molecular weight: 1140.01
化合物21可根据实施例1描述的代表性路线合成。Compound 21 can be synthesized according to the representative route described in Example 1.
H NMR(300MHz,DMSO-d6):1H NMR(DMSO-d6):δ(ppm)5.4(s,5H),3.49(m,5H,),2.22-2.71(m,25H),1.47(m,10H),1.28(m,80H),0.90(m,10H).H NMR(300MHz,DMSO-d6):1H NMR(DMSO-d6):δ(ppm)5.4(s,5H),3.49(m,5H,),2.22-2.71(m,25H),1.47(m, 10H),1.28(m,80H),0.90(m,10H).
实施例23：Example 23:
化合物22的合成Synthesis of compound 22
Figure PCTCN2022084347-appb-000039
Figure PCTCN2022084347-appb-000039
化学式：C 70H 142D 3N 5O 5 Chemical formula: C 70 H 142 D 3 N 5 O 5
分子量：1140.01Molecular weight: 1140.01
化合物22可根据实施例1描述的代表性路线合成。Compound 22 can be synthesized according to the representative route described in Example 1.
H NMR(300MHz,DMSO-d6):1H NMR(DMSO-d6):δ(ppm)5.4(s,5H),3.49(m,5H,),2.22-2.71(m,25H),1.47(m,10H),1.28(m,80H),0.90(m,10H).H NMR(300MHz,DMSO-d6):1H NMR(DMSO-d6):δ(ppm)5.4(s,5H),3.49(m,5H,),2.22-2.71(m,25H),1.47(m, 10H),1.28(m,80H),0.90(m,10H).
实施例24：Example 24:
化合物23的合成Synthesis of compound 23
Figure PCTCN2022084347-appb-000040
Figure PCTCN2022084347-appb-000040
化学式：C 70H 142D 3N 5O 5 Chemical formula: C 70 H 142 D 3 N 5 O 5
分子量：1140.01Molecular weight: 1140.01
化合物23可根据实施例1描述的代表性路线合成。Compound 23 can be synthesized according to the representative route described in Example 1.
H NMR(300MHz,DMSO-d6):1H NMR(DMSO-d6):δ(ppm)5.4(s,5H),3.49(m,5H,),2.22-2.71(m,25H),1.47(m,10H),1.28(m,80H),0.90(m,10H).H NMR(300MHz,DMSO-d6):1H NMR(DMSO-d6):δ(ppm)5.4(s,5H),3.49(m,5H,),2.22-2.71(m,25H),1.47(m, 10H),1.28(m,80H),0.90(m,10H).
实施例25：Example 25:
化合物24的合成Synthesis of Compound 24
Figure PCTCN2022084347-appb-000041
Figure PCTCN2022084347-appb-000041
化学式：C 70H 142D 3N 5O 5 Chemical formula: C 70 H 142 D 3 N 5 O 5
分子量：1140.01Molecular weight: 1140.01
化合物24可根据实施例1描述的代表性路线合成。Compound 24 can be synthesized according to the representative route described in Example 1.
H NMR(300MHz,DMSO-d6):1H NMR(DMSO-d6):δ(ppm)5.4(s,5H),3.49(m,5H,),2.22-2.71(m,25H),1.47(m,10H),1.28(m,80H),0.90(m,10H).H NMR(300MHz,DMSO-d6):1H NMR(DMSO-d6):δ(ppm)5.4(s,5H),3.49(m,5H,),2.22-2.71(m,25H),1.47(m, 10H),1.28(m,80H),0.90(m,10H).
实施例26：Example 26:
化合物25的合成Synthesis of compound 25
Figure PCTCN2022084347-appb-000042
Figure PCTCN2022084347-appb-000042
化学式：C 70H 142D 3N 5O 5 Chemical formula: C 70 H 142 D 3 N 5 O 5
分子量：1140.01Molecular weight: 1140.01
化合物25可根据实施例1描述的代表性路线合成。Compound 25 can be synthesized according to the representative route described in Example 1.
H NMR(300MHz,DMSO-d6):1H NMR(DMSO-d6):δ(ppm)5.4(s,5H),3.49(m,5H,),2.22-2.71(m,25H),1.47(m,10H),1.28(m,80H),0.90(m,10H).H NMR(300MHz,DMSO-d6):1H NMR(DMSO-d6):δ(ppm)5.4(s,5H),3.49(m,5H,),2.22-2.71(m,25H),1.47(m, 10H),1.28(m,80H),0.90(m,10H).
实施例27：Example 27:
化合物26的合成Synthesis of compound 26
Figure PCTCN2022084347-appb-000043
Figure PCTCN2022084347-appb-000043
化学式：C 70H 142D 3N 5O 5 Chemical formula: C 70 H 142 D 3 N 5 O 5
分子量：1140.01Molecular weight: 1140.01
化合物26可根据实施例1描述的代表性路线合成。Compound 26 can be synthesized according to the representative route described in Example 1.
H NMR(300MHz,DMSO-d6):1H NMR(DMSO-d6):δ(ppm)5.4(s,5H),3.49(m,5H,),2.22-2.71(m,25H),1.47(m,10H),1.28(m,80H),0.90(m,10H).H NMR(300MHz,DMSO-d6):1H NMR(DMSO-d6):δ(ppm)5.4(s,5H),3.49(m,5H,),2.22-2.71(m,25H),1.47(m, 10H),1.28(m,80H),0.90(m,10H).
实施例28：Example 28:
化合物27的合成Synthesis of compound 27
Figure PCTCN2022084347-appb-000044
Figure PCTCN2022084347-appb-000044
化学式：C 70H 142D 3N 5O 5 Chemical formula: C 70 H 142 D 3 N 5 O 5
分子量：1140.01Molecular weight: 1140.01
化合物27可根据实施例1描述的代表性路线合成。Compound 27 can be synthesized according to the representative route described in Example 1.
H NMR(300MHz,DMSO-d6):1H NMR(DMSO-d6):δ(ppm)5.4(s,5H),3.49(m,5H,),2.22-2.71(m,24H),1.47(m,10H),1.28(m,80H),0.90(m,10H).H NMR(300MHz,DMSO-d6):1H NMR(DMSO-d6):δ(ppm)5.4(s,5H),3.49(m,5H,),2.22-2.71(m,24H),1.47(m, 10H),1.28(m,80H),0.90(m,10H).
实施例29：Example 29:
化合物28的合成Synthesis of compound 28
Figure PCTCN2022084347-appb-000045
Figure PCTCN2022084347-appb-000045
化学式：C 70H 142D 3N 5O 5 Chemical formula: C 70 H 142 D 3 N 5 O 5
分子量：1140.01Molecular weight: 1140.01
化合物28可根据实施例1描述的代表性路线合成。Compound 28 can be synthesized according to the representative route described in Example 1.
H NMR(300MHz,DMSO-d6):1H NMR(DMSO-d6):δ(ppm)5.4(s,5H),3.49(m,5H,),2.22-2.71(m,25H),1.47(m,10H),1.28(m,80H),0.90(m,10H).H NMR(300MHz,DMSO-d6):1H NMR(DMSO-d6):δ(ppm)5.4(s,5H),3.49(m,5H,),2.22-2.71(m,25H),1.47(m, 10H),1.28(m,80H),0.90(m,10H).
实施例30：Example 30:
化合物29的合成Synthesis of compound 29
Figure PCTCN2022084347-appb-000046
Figure PCTCN2022084347-appb-000046
化学式：C 70H 141D 4N 5O 5 Chemical formula: C 70 H 141 D 4 N 5 O 5
分子量：1141.01Molecular weight: 1141.01
化合物29可根据实施例1描述的代表性路线合成。Compound 29 can be synthesized according to the representative route described in Example 1.
H NMR(300MHz,DMSO-d6):1H NMR(DMSO-d6):δ(ppm)5.4(s,5H),3.49(m,5H,),2.22-2.71(m,24H),1.47(m,10H),1.28(m,80H),0.90(m,10H).H NMR(300MHz,DMSO-d6):1H NMR(DMSO-d6):δ(ppm)5.4(s,5H),3.49(m,5H,),2.22-2.71(m,24H),1.47(m, 10H),1.28(m,80H),0.90(m,10H).
实施例31：Example 31:
化合物30的合成Synthesis of compound 30
Figure PCTCN2022084347-appb-000047
Figure PCTCN2022084347-appb-000047
化学式：C 70H 141D 4N 5O 5 Chemical formula: C 70 H 141 D 4 N 5 O 5
分子量：1141.01Molecular weight: 1141.01
化合物30可根据实施例1描述的代表性路线合成。Compound 30 can be synthesized according to the representative route described in Example 1.
H NMR(300MHz,DMSO-d6):1H NMR(DMSO-d6):δ(ppm)5.4(s,5H),3.49(m,5H,),2.22-2.71(m,24H),1.47(m,10H),1.28(m,80H),0.90(m,10H).H NMR(300MHz,DMSO-d6):1H NMR(DMSO-d6):δ(ppm)5.4(s,5H),3.49(m,5H,),2.22-2.71(m,24H),1.47(m, 10H),1.28(m,80H),0.90(m,10H).
实施例32：Example 32:
化合物31的合成Synthesis of compound 31
Figure PCTCN2022084347-appb-000048
Figure PCTCN2022084347-appb-000048
化学式：C 70H 141D 4N 5O 5 Chemical formula: C 70 H 141 D 4 N 5 O 5
分子量：1141.01Molecular weight: 1141.01
化合物31可根据实施例1描述的代表性路线合成。Compound 31 can be synthesized according to the representative route described in Example 1.
H NMR(300MHz,DMSO-d6):1H NMR(DMSO-d6):δ(ppm)5.4(s,5H),3.49(m,5H,),2.22-2.71(m,24H),1.47(m,10H),1.28(m,80H),0.90(m,10H).H NMR(300MHz,DMSO-d6):1H NMR(DMSO-d6):δ(ppm)5.4(s,5H),3.49(m,5H,),2.22-2.71(m,24H),1.47(m, 10H),1.28(m,80H),0.90(m,10H).
实施例33：Example 33:
化合物32的合成Synthesis of compound 32
Figure PCTCN2022084347-appb-000049
Figure PCTCN2022084347-appb-000049
化学式：C 70H 141D 4N 5O 5 Chemical formula: C 70 H 141 D 4 N 5 O 5
分子量：1141.01Molecular weight: 1141.01
化合物32可根据实施例1描述的代表性路线合成。Compound 32 can be synthesized according to the representative route described in Example 1.
H NMR(300MHz,DMSO-d6):1H NMR(DMSO-d6):δ(ppm)5.4(s,5H),3.49(m,5H,),2.22-2.71(m,24H),1.47(m,10H),1.28(m,80H),0.90(m,10H).H NMR(300MHz,DMSO-d6):1H NMR(DMSO-d6):δ(ppm)5.4(s,5H),3.49(m,5H,),2.22-2.71(m,24H),1.47(m, 10H),1.28(m,80H),0.90(m,10H).
实施例34：Example 34:
化合物33的合成Synthesis of Compound 33
Figure PCTCN2022084347-appb-000050
Figure PCTCN2022084347-appb-000050
化学式：C 70H 141D 4N 5O 5 Chemical formula: C 70 H 141 D 4 N 5 O 5
分子量：1141.01Molecular weight: 1141.01
化合物33可根据实施例1描述的代表性路线合成。Compound 33 can be synthesized according to the representative route described in Example 1.
H NMR(300MHz,DMSO-d6):1H NMR(DMSO-d6):δ(ppm)5.4(s,5H),3.49(m,5H,),2.22-2.71(m,24H),1.47(m,10H),1.28(m,80H),0.90(m,10H).H NMR(300MHz,DMSO-d6):1H NMR(DMSO-d6):δ(ppm)5.4(s,5H),3.49(m,5H,),2.22-2.71(m,24H),1.47(m, 10H),1.28(m,80H),0.90(m,10H).
实施例35：Example 35:
化合物34的合成Synthesis of compound 34
Figure PCTCN2022084347-appb-000051
Figure PCTCN2022084347-appb-000051
化学式：C 70H 141D 4N 5O 5 Chemical formula: C 70 H 141 D 4 N 5 O 5
分子量：1141.01Molecular weight: 1141.01
化合物34可根据实施例1描述的代表性路线合成。Compound 34 can be synthesized according to the representative route described in Example 1.
H NMR(300MHz,DMSO-d6):1H NMR(DMSO-d6):δ(ppm)5.4(s,5H),3.49(m,5H,),2.22-2.71(m,24H),1.47(m,10H),1.28(m,80H),0.90(m,10H).H NMR(300MHz,DMSO-d6):1H NMR(DMSO-d6):δ(ppm)5.4(s,5H),3.49(m,5H,),2.22-2.71(m,24H),1.47(m, 10H),1.28(m,80H),0.90(m,10H).
实施例36：Example 36:
化合物35的合成Synthesis of compound 35
Figure PCTCN2022084347-appb-000052
Figure PCTCN2022084347-appb-000052
化学式：C 70H 141D 4N 5O 5 Chemical formula: C 70 H 141 D 4 N 5 O 5
分子量：1141.01Molecular weight: 1141.01
化合物35可根据实施例1描述的代表性路线合成。Compound 35 can be synthesized according to the representative route described in Example 1.
H NMR(300MHz,DMSO-d6):1H NMR(DMSO-d6):δ(ppm)5.4(s,5H),3.49(m,5H,),2.22-2.71(m,25H),1.47(m,10H),1.28(m,80H),0.90(m,10H).H NMR(300MHz,DMSO-d6):1H NMR(DMSO-d6):δ(ppm)5.4(s,5H),3.49(m,5H,),2.22-2.71(m,25H),1.47(m, 10H),1.28(m,80H),0.90(m,10H).
实施例37：Example 37:
化合物36的合成Synthesis of Compound 36
Figure PCTCN2022084347-appb-000053
Figure PCTCN2022084347-appb-000053
化学式：C 70H 141D 4N 5O 5 Chemical formula: C 70 H 141 D 4 N 5 O 5
分子量：1141.01Molecular weight: 1141.01
化合物36可根据实施例1描述的代表性路线合成。Compound 36 can be synthesized according to the representative route described in Example 1.
H NMR(300MHz,DMSO-d6):1H NMR(DMSO-d6):δ(ppm)5.4(s,5H),3.49(m,5H,),2.22-2.71(m,24H),1.47(m,10H),1.28(m,80H),0.90(m,10H).H NMR(300MHz,DMSO-d6):1H NMR(DMSO-d6):δ(ppm)5.4(s,5H),3.49(m,5H,),2.22-2.71(m,24H),1.47(m, 10H),1.28(m,80H),0.90(m,10H).
实施例38：Example 38:
化合物37的合成Synthesis of Compound 37
Figure PCTCN2022084347-appb-000054
Figure PCTCN2022084347-appb-000054
化学式：C 70H 141D 4N 5O 5 Chemical formula: C 70 H 141 D 4 N 5 O 5
分子量：1141.01Molecular weight: 1141.01
化合物37可根据实施例1描述的代表性路线合成。Compound 37 can be synthesized according to the representative route described in Example 1.
H NMR(300MHz,DMSO-d6):1H NMR(DMSO-d6):δ(ppm)5.4(s,5H),3.49(m,5H,),2.22-2.71(m,24H),1.47(m,10H),1.28(m,80H),0.90(m,10H).H NMR(300MHz,DMSO-d6):1H NMR(DMSO-d6):δ(ppm)5.4(s,5H),3.49(m,5H,),2.22-2.71(m,24H),1.47(m, 10H),1.28(m,80H),0.90(m,10H).
实施例39：Example 39:
化合物38的合成Synthesis of compound 38
Figure PCTCN2022084347-appb-000055
Figure PCTCN2022084347-appb-000055
化学式：C 70H 141D 4N 5O 5 Chemical formula: C 70 H 141 D 4 N 5 O 5
分子量：1141.01Molecular weight: 1141.01
化合物38可根据实施例1描述的代表性路线合成。Compound 38 can be synthesized according to the representative route described in Example 1.
H NMR(300MHz,DMSO-d6):1H NMR(DMSO-d6):δ(ppm)5.4(s,5H),3.49(m,5H,),2.22-2.71(m,24H),1.47(m,10H),1.28(m,80H),0.90(m,10H).H NMR(300MHz,DMSO-d6):1H NMR(DMSO-d6):δ(ppm)5.4(s,5H),3.49(m,5H,),2.22-2.71(m,24H),1.47(m, 10H),1.28(m,80H),0.90(m,10H).
实施例40：Example 40:
化合物39的合成Synthesis of compound 39
Figure PCTCN2022084347-appb-000056
Figure PCTCN2022084347-appb-000056
化学式：C 70H 141D 4N 5O 5 Chemical formula: C 70 H 141 D 4 N 5 O 5
分子量：1141.01Molecular weight: 1141.01
化合物39可根据实施例1描述的代表性路线合成。Compound 39 can be synthesized according to the representative route described in Example 1.
H NMR(300MHz,DMSO-d6):1H NMR(DMSO-d6):δ(ppm)5.4(s,5H),3.49(m,5H,),2.22-2.71(m,24H),1.47(m,10H),1.28(m,80H),0.90(m,10H).H NMR(300MHz,DMSO-d6):1H NMR(DMSO-d6):δ(ppm)5.4(s,5H),3.49(m,5H,),2.22-2.71(m,24H),1.47(m, 10H),1.28(m,80H),0.90(m,10H).
实施例41：Example 41:
化合物40的合成Synthesis of Compound 40
Figure PCTCN2022084347-appb-000057
Figure PCTCN2022084347-appb-000057
化学式：C 70H 141D 4N 5O 5 Chemical formula: C 70 H 141 D 4 N 5 O 5
分子量：1141.01Molecular weight: 1141.01
化合物40可根据实施例1描述的代表性路线合成。Compound 40 can be synthesized according to the representative route described in Example 1.
H NMR(300MHz,DMSO-d6):1H NMR(DMSO-d6):δ(ppm)5.4(s,5H),3.49(m,5H,),2.22-2.71(m,24H),1.47(m,10H),1.28(m,80H),0.90(m,10H).H NMR(300MHz,DMSO-d6):1H NMR(DMSO-d6):δ(ppm)5.4(s,5H),3.49(m,5H,),2.22-2.71(m,24H),1.47(m, 10H),1.28(m,80H),0.90(m,10H).
实施例42：Example 42:
化合物41的合成Synthesis of Compound 41
Figure PCTCN2022084347-appb-000058
Figure PCTCN2022084347-appb-000058
化学式：C 70H 141D 4N 5O 5 Chemical formula: C 70 H 141 D 4 N 5 O 5
分子量：1141.01Molecular weight: 1141.01
化合物41可根据实施例1描述的代表性路线合成。Compound 41 can be synthesized according to the representative route described in Example 1.
H NMR(300MHz,DMSO-d6):1H NMR(DMSO-d6):δ(ppm)5.4(s,5H),3.49(m,5H,),2.22-2.71(m,24H),1.47(m,10H),1.28(m,80H),0.90(m,10H).H NMR(300MHz,DMSO-d6):1H NMR(DMSO-d6):δ(ppm)5.4(s,5H),3.49(m,5H,),2.22-2.71(m,24H),1.47(m, 10H),1.28(m,80H),0.90(m,10H).
实施例43：Example 43:
化合物42的合成Synthesis of compound 42
Figure PCTCN2022084347-appb-000059
Figure PCTCN2022084347-appb-000059
化学式：C 70H 141D 4N 5O 5 Chemical formula: C 70 H 141 D 4 N 5 O 5
分子量：1141.01Molecular weight: 1141.01
化合物42可根据实施例1描述的代表性路线合成。Compound 42 can be synthesized according to the representative route described in Example 1.
H NMR(300MHz,DMSO-d6):1H NMR(DMSO-d6):δ(ppm)5.4(s,5H),3.49(m,5H,),2.22-2.71(m,24H),1.47(m,10H),1.28(m,80H),0.90(m,10H).H NMR(300MHz,DMSO-d6):1H NMR(DMSO-d6):δ(ppm)5.4(s,5H),3.49(m,5H,),2.22-2.71(m,24H),1.47(m, 10H),1.28(m,80H),0.90(m,10H).
实施例44：Example 44:
化合物43的合成Synthesis of Compound 43
Figure PCTCN2022084347-appb-000060
Figure PCTCN2022084347-appb-000060
化学式：C 70H 141D 4N 5O 5 Chemical formula: C 70 H 141 D 4 N 5 O 5
分子量：1141.01Molecular weight: 1141.01
化合物43可根据实施例1描述的代表性路线合成。Compound 43 can be synthesized according to the representative route described in Example 1.
H NMR(300MHz,DMSO-d6):1H NMR(DMSO-d6):δ(ppm)5.4(s,5H),3.49(m,5H,),2.22-2.71(m,24H),1.47(m,10H),1.28(m,80H),0.90(m,10H).H NMR(300MHz,DMSO-d6):1H NMR(DMSO-d6):δ(ppm)5.4(s,5H),3.49(m,5H,),2.22-2.71(m,24H),1.47(m, 10H),1.28(m,80H),0.90(m,10H).
实施例45：Example 45:
化合物44的合成Synthesis of compound 44
Figure PCTCN2022084347-appb-000061
Figure PCTCN2022084347-appb-000061
化学式：C 70H 141D 4N 5O 5 Chemical formula: C 70 H 141 D 4 N 5 O 5
分子量：1141.01Molecular weight: 1141.01
化合物44可根据实施例1描述的代表性路线合成。Compound 44 can be synthesized according to the representative route described in Example 1.
H NMR(300MHz,DMSO-d6):1H NMR(DMSO-d6):δ(ppm)5.4(s,5H),3.49(m,5H,),2.22-2.71(m,24H),1.47(m,10H),1.28(m,80H),0.90(m,10H).H NMR(300MHz,DMSO-d6):1H NMR(DMSO-d6):δ(ppm)5.4(s,5H),3.49(m,5H,),2.22-2.71(m,24H),1.47(m, 10H),1.28(m,80H),0.90(m,10H).
实施例46：Example 46:
化合物45的合成Synthesis of Compound 45
Figure PCTCN2022084347-appb-000062
Figure PCTCN2022084347-appb-000062
化学式：C 70H 140D 5N 5O 5 Chemical formula: C 70 H 140 D 5 N 5 O 5
分子量：1142.01Molecular weight: 1142.01
化合物45可根据实施例1描述的代表性路线合成。Compound 45 can be synthesized according to the representative route described in Example 1.
H NMR(300MHz,DMSO-d6):1H NMR(DMSO-d6):δ(ppm)5.4(s,5H),3.49(m,5H,),2.22-2.71(m,23H),1.47(m,10H),1.28(m,80H),0.90(m,10H).H NMR(300MHz,DMSO-d6):1H NMR(DMSO-d6):δ(ppm)5.4(s,5H),3.49(m,5H,),2.22-2.71(m,23H),1.47(m, 10H),1.28(m,80H),0.90(m,10H).
实施例47：Example 47:
化合物46的合成Synthesis of compound 46
Figure PCTCN2022084347-appb-000063
Figure PCTCN2022084347-appb-000063
化学式：C 70H 140D 5N 5O 5 Chemical formula: C 70 H 140 D 5 N 5 O 5
分子量：1142.01Molecular weight: 1142.01
化合物46可根据实施例1描述的代表性路线合成。Compound 46 can be synthesized according to the representative route described in Example 1.
H NMR(300MHz,DMSO-d6):1H NMR(DMSO-d6):δ(ppm)5.4(s,5H),3.49(m,5H,),2.22-2.71(m,23H),1.47(m,10H),1.28(m,80H),0.90(m,10H).H NMR(300MHz,DMSO-d6):1H NMR(DMSO-d6):δ(ppm)5.4(s,5H),3.49(m,5H,),2.22-2.71(m,23H),1.47(m, 10H),1.28(m,80H),0.90(m,10H).
实施例48：Example 48:
化合物47的合成Synthesis of compound 47
Figure PCTCN2022084347-appb-000064
Figure PCTCN2022084347-appb-000064
化学式：C 70H 140D 5N 5O 5 Chemical formula: C 70 H 140 D 5 N 5 O 5
分子量：1142.01Molecular weight: 1142.01
化合物47可根据实施例1描述的代表性路线合成。Compound 47 can be synthesized according to the representative route described in Example 1.
H NMR(300MHz,DMSO-d6):1H NMR(DMSO-d6):δ(ppm)5.4(s,5H),3.49(m,5H,),2.22-2.71(m,23H),1.47(m,10H),1.28(m,80H),0.90(m,10H).H NMR(300MHz,DMSO-d6):1H NMR(DMSO-d6):δ(ppm)5.4(s,5H),3.49(m,5H,),2.22-2.71(m,23H),1.47(m, 10H),1.28(m,80H),0.90(m,10H).
实施例49：Example 49:
化合物48的合成Synthesis of Compound 48
Figure PCTCN2022084347-appb-000065
Figure PCTCN2022084347-appb-000065
化学式：C 70H 140D 5N 5O 5 Chemical formula: C 70 H 140 D 5 N 5 O 5
分子量：1142.01Molecular weight: 1142.01
化合物48可根据实施例1描述的代表性路线合成。Compound 48 can be synthesized according to the representative route described in Example 1.
H NMR(300MHz,DMSO-d6):1H NMR(DMSO-d6):δ(ppm)5.4(s,5H),3.49(m,5H,),2.22-2.71(m,23H),1.47(m,10H),1.28(m,80H),0.90(m,10H).H NMR(300MHz,DMSO-d6):1H NMR(DMSO-d6):δ(ppm)5.4(s,5H),3.49(m,5H,),2.22-2.71(m,23H),1.47(m, 10H),1.28(m,80H),0.90(m,10H).
实施例50：Example 50:
化合物49的合成Synthesis of Compound 49
Figure PCTCN2022084347-appb-000066
Figure PCTCN2022084347-appb-000066
化学式：C 70H 140D 5N 5O 5 Chemical formula: C 70 H 140 D 5 N 5 O 5
分子量：1142.01Molecular weight: 1142.01
化合物49可根据实施例1描述的代表性路线合成。Compound 49 can be synthesized according to the representative route described in Example 1.
H NMR(300MHz,DMSO-d6):1H NMR(DMSO-d6):δ(ppm)5.4(s,5H),3.49(m,5H,),2.22-2.71(m,23H),1.47(m,10H),1.28(m,80H),0.90(m,10H).H NMR(300MHz,DMSO-d6):1H NMR(DMSO-d6):δ(ppm)5.4(s,5H),3.49(m,5H,),2.22-2.71(m,23H),1.47(m, 10H),1.28(m,80H),0.90(m,10H).
实施例51：Example 51:
化合物50的合成Synthesis of Compound 50
Figure PCTCN2022084347-appb-000067
Figure PCTCN2022084347-appb-000067
化学式：C 70H 140D 5N 5O 5 Chemical formula: C 70 H 140 D 5 N 5 O 5
分子量：1142.01Molecular weight: 1142.01
化合物50可根据实施例1描述的代表性路线合成。Compound 50 can be synthesized according to the representative route described in Example 1.
H NMR(300MHz,DMSO-d6):1H NMR(DMSO-d6):δ(ppm)5.4(s,5H),3.49(m,5H,),2.22-2.71(m,23H),1.47(m,10H),1.28(m,80H),0.90(m,10H).H NMR(300MHz,DMSO-d6):1H NMR(DMSO-d6):δ(ppm)5.4(s,5H),3.49(m,5H,),2.22-2.71(m,23H),1.47(m, 10H),1.28(m,80H),0.90(m,10H).
实施例52：Example 52:
化合物51的合成Synthesis of compound 51
Figure PCTCN2022084347-appb-000068
Figure PCTCN2022084347-appb-000068
化学式：C 70H 140D 5N 5O 5 Chemical formula: C 70 H 140 D 5 N 5 O 5
分子量：1142.01Molecular weight: 1142.01
化合物51可根据实施例1描述的代表性路线合成。Compound 51 can be synthesized according to the representative route described in Example 1.
H NMR(300MHz,DMSO-d6):1H NMR(DMSO-d6):δ(ppm)5.4(s,5H),3.49(m,5H,),2.22-2.71(m,23H),1.47(m,10H),1.28(m,80H),0.90(m,10H).H NMR(300MHz,DMSO-d6):1H NMR(DMSO-d6):δ(ppm)5.4(s,5H),3.49(m,5H,),2.22-2.71(m,23H),1.47(m, 10H),1.28(m,80H),0.90(m,10H).
实施例53：Example 53:
化合物52的合成Synthesis of compound 52
Figure PCTCN2022084347-appb-000069
Figure PCTCN2022084347-appb-000069
化学式：C 70H 140D 5N 5O 5 Chemical formula: C 70 H 140 D 5 N 5 O 5
分子量：1142.01Molecular weight: 1142.01
化合物53可根据实施例1描述的代表性路线合成。Compound 53 can be synthesized according to the representative route described in Example 1.
H NMR(300MHz,DMSO-d6):1H NMR(DMSO-d6):δ(ppm)5.4(s,5H),3.49(m,5H,),2.22-2.71(m,24H),1.47(m,10H),1.28(m,80H),0.90(m,10H).H NMR(300MHz,DMSO-d6):1H NMR(DMSO-d6):δ(ppm)5.4(s,5H),3.49(m,5H,),2.22-2.71(m,24H),1.47(m, 10H),1.28(m,80H),0.90(m,10H).
实施例54：Example 54:
化合物53的合成Synthesis of compound 53
Figure PCTCN2022084347-appb-000070
Figure PCTCN2022084347-appb-000070
化学式：C 70H 140D 5N 5O 5 Chemical formula: C 70 H 140 D 5 N 5 O 5
分子量：1142.01Molecular weight: 1142.01
化合物53可根据实施例1描述的代表性路线合成。Compound 53 can be synthesized according to the representative route described in Example 1.
H NMR(300MHz,DMSO-d6):1H NMR(DMSO-d6):δ(ppm)5.4(s,5H),3.49(m,5H,),2.22-2.71(m,23H),1.47(m,10H),1.28(m,80H),0.90(m,10H).H NMR(300MHz,DMSO-d6):1H NMR(DMSO-d6):δ(ppm)5.4(s,5H),3.49(m,5H,),2.22-2.71(m,23H),1.47(m, 10H),1.28(m,80H),0.90(m,10H).
实施例55：Example 55:
化合物54的合成Synthesis of Compound 54
Figure PCTCN2022084347-appb-000071
Figure PCTCN2022084347-appb-000071
化学式：C 70H 140D 5N 5O 5 Chemical formula: C 70 H 140 D 5 N 5 O 5
分子量：1142.01Molecular weight: 1142.01
化合物54可根据实施例1描述的代表性路线合成。Compound 54 can be synthesized according to the representative route described in Example 1.
H NMR(300MHz,DMSO-d6):1H NMR(DMSO-d6):δ(ppm)5.4(s,5H),3.49(m,5H,),2.22-2.71(m,23H),1.47(m,10H),1.28(m,80H),0.90(m,10H).H NMR(300MHz,DMSO-d6):1H NMR(DMSO-d6):δ(ppm)5.4(s,5H),3.49(m,5H,),2.22-2.71(m,23H),1.47(m, 10H),1.28(m,80H),0.90(m,10H).
实施例56：Example 56:
化合物55的合成Synthesis of Compound 55
Figure PCTCN2022084347-appb-000072
Figure PCTCN2022084347-appb-000072
化学式：C 70H 140D 5N 5O 5 Chemical formula: C 70 H 140 D 5 N 5 O 5
分子量：1142.01Molecular weight: 1142.01
化合物55可根据实施例1描述的代表性路线合成。Compound 55 can be synthesized according to the representative route described in Example 1.
H NMR(300MHz,DMSO-d6):1H NMR(DMSO-d6):δ(ppm)5.4(s,5H),3.49(m,5H,),2.22-2.71(m,23H),1.47(m,10H),1.28(m,80H),0.90(m,10H).H NMR(300MHz,DMSO-d6):1H NMR(DMSO-d6):δ(ppm)5.4(s,5H),3.49(m,5H,),2.22-2.71(m,23H),1.47(m, 10H),1.28(m,80H),0.90(m,10H).
实施例57：Example 57:
化合物56的合成Synthesis of compound 56
Figure PCTCN2022084347-appb-000073
Figure PCTCN2022084347-appb-000073
化学式：C 70H 140D 5N 5O 5 Chemical formula: C 70 H 140 D 5 N 5 O 5
分子量：1142.01Molecular weight: 1142.01
化合物56可根据实施例1描述的代表性路线合成。Compound 56 can be synthesized according to the representative route described in Example 1.
H NMR(300MHz,DMSO-d6):1H NMR(DMSO-d6):δ(ppm)5.4(s,5H),3.49(m,5H,),2.22-2.71(m,23H),1.47(m,10H),1.28(m,80H),0.90(m,10H).H NMR(300MHz,DMSO-d6):1H NMR(DMSO-d6):δ(ppm)5.4(s,5H),3.49(m,5H,),2.22-2.71(m,23H),1.47(m, 10H),1.28(m,80H),0.90(m,10H).
实施例58：Example 58:
化合物57的合成Synthesis of compound 57
Figure PCTCN2022084347-appb-000074
Figure PCTCN2022084347-appb-000074
化学式：C 70H 140D 5N 5O 5 Chemical formula: C 70 H 140 D 5 N 5 O 5
分子量：1142.01Molecular weight: 1142.01
化合物57可根据实施例1描述的代表性路线合成。Compound 57 can be synthesized according to the representative route described in Example 1.
H NMR(300MHz,DMSO-d6):1H NMR(DMSO-d6):δ(ppm)5.4(s,5H),3.49(m,5H,),2.22-2.71(m,23H),1.47(m,10H),1.28(m,80H),0.90(m,10H).H NMR(300MHz,DMSO-d6):1H NMR(DMSO-d6):δ(ppm)5.4(s,5H),3.49(m,5H,),2.22-2.71(m,23H),1.47(m, 10H),1.28(m,80H),0.90(m,10H).
实施例59：Example 59:
化合物58的合成Synthesis of Compound 58
Figure PCTCN2022084347-appb-000075
Figure PCTCN2022084347-appb-000075
化学式：C 70H 140D 5N 5O 5 Chemical formula: C 70 H 140 D 5 N 5 O 5
分子量：1142.01Molecular weight: 1142.01
化合物58可根据实施例1描述的代表性路线合成。Compound 58 can be synthesized according to the representative route described in Example 1.
H NMR(300MHz,DMSO-d6):1H NMR(DMSO-d6):δ(ppm)5.4(s,5H),3.49(m,5H,),2.22-2.71(m,23H),1.47(m,10H),1.28(m,80H),0.90(m,10H).H NMR(300MHz,DMSO-d6):1H NMR(DMSO-d6):δ(ppm)5.4(s,5H),3.49(m,5H,),2.22-2.71(m,23H),1.47(m, 10H),1.28(m,80H),0.90(m,10H).
实施例60：Example 60:
化合物59的合成Synthesis of Compound 59
Figure PCTCN2022084347-appb-000076
Figure PCTCN2022084347-appb-000076
化学式：C 70H 140D 5N 5O 5 Chemical formula: C 70 H 140 D 5 N 5 O 5
分子量：1142.01Molecular weight: 1142.01
化合物59可根据实施例1描述的代表性路线合成。Compound 59 can be synthesized according to the representative route described in Example 1.
H NMR(300MHz,DMSO-d6):1H NMR(DMSO-d6):δ(ppm)5.4(s,5H),3.49(m,5H,),2.22-2.71(m,23H),1.47(m,10H),1.28(m,80H),0.90(m,10H).H NMR(300MHz,DMSO-d6):1H NMR(DMSO-d6):δ(ppm)5.4(s,5H),3.49(m,5H,),2.22-2.71(m,23H),1.47(m, 10H),1.28(m,80H),0.90(m,10H).
实施例61：Example 61:
化合物60的合成Synthesis of Compound 60
Figure PCTCN2022084347-appb-000077
Figure PCTCN2022084347-appb-000077
化学式：C 70H 140D 5N 5O 5 Chemical formula: C 70 H 140 D 5 N 5 O 5
分子量：1142.01Molecular weight: 1142.01
化合物60可根据实施例1描述的代表性路线合成。Compound 60 can be synthesized according to the representative route described in Example 1.
H NMR(300MHz,DMSO-d6):1H NMR(DMSO-d6):δ(ppm)5.4(s,5H),3.49(m,5H,),2.22-2.71(m,23H),1.47(m,10H),1.28(m,80H),0.90(m,10H).H NMR(300MHz,DMSO-d6):1H NMR(DMSO-d6):δ(ppm)5.4(s,5H),3.49(m,5H,),2.22-2.71(m,23H),1.47(m, 10H),1.28(m,80H),0.90(m,10H).
实施例62：Example 62:
化合物61的合成Synthesis of Compound 61
Figure PCTCN2022084347-appb-000078
Figure PCTCN2022084347-appb-000078
化学式：C 70H 139D 6N 5O 5 Chemical formula: C 70 H 139 D 6 N 5 O 5
分子量：1143.01Molecular weight: 1143.01
化合物61可根据实施例1描述的代表性路线合成。Compound 61 can be synthesized according to the representative route described in Example 1.
H NMR(300MHz,DMSO-d6):1H NMR(DMSO-d6):δ(ppm)5.4(s,5H),3.49(m,5H,),2.22-2.71(m,22H),1.47(m,10H),1.28(m,80H),0.90(m,10H).H NMR(300MHz,DMSO-d6):1H NMR(DMSO-d6):δ(ppm)5.4(s,5H),3.49(m,5H,),2.22-2.71(m,22H),1.47(m, 10H),1.28(m,80H),0.90(m,10H).
实施例63：Example 63:
化合物62的合成Synthesis of compound 62
Figure PCTCN2022084347-appb-000079
Figure PCTCN2022084347-appb-000079
化学式：C 70H 139D 6N 5O 5 Chemical formula: C 70 H 139 D 6 N 5 O 5
分子量：1143.01Molecular weight: 1143.01
化合物62可根据实施例1描述的代表性路线合成。Compound 62 can be synthesized according to the representative route described in Example 1.
H NMR(300MHz,DMSO-d6):1H NMR(DMSO-d6):δ(ppm)5.4(s,5H),3.49(m,5H,),2.22-2.71(m,22H),1.47(m,10H),1.28(m,80H),0.90(m,10H).H NMR(300MHz,DMSO-d6):1H NMR(DMSO-d6):δ(ppm)5.4(s,5H),3.49(m,5H,),2.22-2.71(m,22H),1.47(m, 10H),1.28(m,80H),0.90(m,10H).
实施例64：Example 64:
化合物63的合成Synthesis of Compound 63
Figure PCTCN2022084347-appb-000080
Figure PCTCN2022084347-appb-000080
化学式：C 70H 139D 6N 5O 5 Chemical formula: C 70 H 139 D 6 N 5 O 5
分子量：1143.01Molecular weight: 1143.01
化合物63可根据实施例1描述的代表性路线合成。Compound 63 can be synthesized according to the representative route described in Example 1.
H NMR(300MHz,DMSO-d6):1H NMR(DMSO-d6):δ(ppm)5.4(s,5H),3.49(m,5H,),2.22-2.71(m,22H),1.47(m,10H),1.28(m,80H),0.90(m,10H).H NMR(300MHz,DMSO-d6):1H NMR(DMSO-d6):δ(ppm)5.4(s,5H),3.49(m,5H,),2.22-2.71(m,22H),1.47(m, 10H),1.28(m,80H),0.90(m,10H).
实施例65：Example 65:
化合物64的合成Synthesis of Compound 64
Figure PCTCN2022084347-appb-000081
Figure PCTCN2022084347-appb-000081
化学式：C 70H 139D 6N 5O 5 Chemical formula: C 70 H 139 D 6 N 5 O 5
分子量：1143.01Molecular weight: 1143.01
化合物64可根据实施例1描述的代表性路线合成。Compound 64 can be synthesized according to the representative route described in Example 1.
H NMR(300MHz,DMSO-d6):1H NMR(DMSO-d6):δ(ppm)5.4(s,5H),3.49(m,5H,),2.22-2.71(m,22H),1.47(m,10H),1.28(m,80H),0.90(m,10H).H NMR(300MHz,DMSO-d6):1H NMR(DMSO-d6):δ(ppm)5.4(s,5H),3.49(m,5H,),2.22-2.71(m,22H),1.47(m, 10H),1.28(m,80H),0.90(m,10H).
实施例66：Example 66:
化合物65的合成Synthesis of compound 65
Figure PCTCN2022084347-appb-000082
Figure PCTCN2022084347-appb-000082
化学式：C 70H 139D 6N 5O 5 Chemical formula: C 70 H 139 D 6 N 5 O 5
分子量：1143.01Molecular weight: 1143.01
化合物65可根据实施例1描述的代表性路线合成。Compound 65 can be synthesized according to the representative route described in Example 1.
H NMR(300MHz,DMSO-d6):1H NMR(DMSO-d6):δ(ppm)5.4(s,5H),3.49(m,5H,),2.22-2.71(m,22H),1.47(m,10H),1.28(m,80H),0.90(m,10H).H NMR(300MHz,DMSO-d6):1H NMR(DMSO-d6):δ(ppm)5.4(s,5H),3.49(m,5H,),2.22-2.71(m,22H),1.47(m, 10H),1.28(m,80H),0.90(m,10H).
实施例67：Example 67:
化合物66的合成Synthesis of Compound 66
Figure PCTCN2022084347-appb-000083
Figure PCTCN2022084347-appb-000083
化学式：C 70H 139D 6N 5O 5 Chemical formula: C 70 H 139 D 6 N 5 O 5
分子量：1143.01Molecular weight: 1143.01
化合物66可根据实施例1描述的代表性路线合成。Compound 66 can be synthesized according to the representative route described in Example 1.
H NMR(300MHz,DMSO-d6):1H NMR(DMSO-d6):δ(ppm)5.4(s,5H),3.49(m,5H,),2.22-2.71(m,22H),1.47(m,10H),1.28(m,80H),0.90(m,10H).H NMR(300MHz,DMSO-d6):1H NMR(DMSO-d6):δ(ppm)5.4(s,5H),3.49(m,5H,),2.22-2.71(m,22H),1.47(m, 10H),1.28(m,80H),0.90(m,10H).
实施例68：Example 68:
化合物67的合成Synthesis of Compound 67
Figure PCTCN2022084347-appb-000084
Figure PCTCN2022084347-appb-000084
化学式：C 70H 139D 6N 5O 5 Chemical formula: C 70 H 139 D 6 N 5 O 5
分子量：1143.01Molecular weight: 1143.01
物67可根据实施例1描述的代表性路线合成。Compound 67 can be synthesized according to the representative route described in Example 1.
H NMR(300MHz,DMSO-d6):1H NMR(DMSO-d6):δ(ppm)5.4(s,5H),3.49(m,5H,),2.22-2.71(m,22H),1.47(m,10H),1.28(m,80H),0.90(m,10H).H NMR(300MHz,DMSO-d6):1H NMR(DMSO-d6):δ(ppm)5.4(s,5H),3.49(m,5H,),2.22-2.71(m,22H),1.47(m, 10H),1.28(m,80H),0.90(m,10H).
实施例69：Example 69:
化合物68的合成Synthesis of Compound 68
Figure PCTCN2022084347-appb-000085
Figure PCTCN2022084347-appb-000085
化学式：C 70H 139D 6N 5O 5 Chemical formula: C 70 H 139 D 6 N 5 O 5
分子量：1143.01Molecular weight: 1143.01
化合物68可根据实施例1描述的代表性路线合成。Compound 68 can be synthesized according to the representative route described in Example 1.
H NMR(300MHz,DMSO-d6):1H NMR(DMSO-d6):δ(ppm)5.4(s,5H),3.49(m,5H,),2.22-2.71(m,22H),1.47(m,10H),1.28(m,80H),0.90(m,10H).H NMR(300MHz,DMSO-d6):1H NMR(DMSO-d6):δ(ppm)5.4(s,5H),3.49(m,5H,),2.22-2.71(m,22H),1.47(m, 10H),1.28(m,80H),0.90(m,10H).
实施例70：Example 70:
化合物69的合成Synthesis of Compound 69
Figure PCTCN2022084347-appb-000086
Figure PCTCN2022084347-appb-000086
化学式：C 70H 139D 6N 5O 5 Chemical formula: C 70 H 139 D 6 N 5 O 5
分子量：1143.01Molecular weight: 1143.01
化合物69可根据实施例1描述的代表性路线合成。Compound 69 can be synthesized according to the representative route described in Example 1.
H NMR(300MHz,DMSO-d6):1H NMR(DMSO-d6):δ(ppm)5.4(s,5H),3.49(m,5H,),2.22-2.71(m,22H),1.47(m,10H),1.28(m,80H),0.90(m,10H).H NMR(300MHz,DMSO-d6):1H NMR(DMSO-d6):δ(ppm)5.4(s,5H),3.49(m,5H,),2.22-2.71(m,22H),1.47(m, 10H),1.28(m,80H),0.90(m,10H).
实施例71：Example 71:
化合物70的合成Synthesis of Compound 70
Figure PCTCN2022084347-appb-000087
Figure PCTCN2022084347-appb-000087
化学式：C 70H 139D 6N 5O 5 Chemical formula: C 70 H 139 D 6 N 5 O 5
分子量：1143.01Molecular weight: 1143.01
化合物70可根据实施例1描述的代表性路线合成。Compound 70 can be synthesized according to the representative route described in Example 1.
H NMR(300MHz,DMSO-d6):1H NMR(DMSO-d6):δ(ppm)5.4(s,5H),3.49(m,5H,),2.22-2.71(m,22H),1.47(m,10H),1.28(m,80H),0.90(m,10H).H NMR(300MHz,DMSO-d6):1H NMR(DMSO-d6):δ(ppm)5.4(s,5H),3.49(m,5H,),2.22-2.71(m,22H),1.47(m, 10H),1.28(m,80H),0.90(m,10H).
实施例72：Example 72:
化合物71的合成Synthesis of compound 71
Figure PCTCN2022084347-appb-000088
Figure PCTCN2022084347-appb-000088
化学式：C 70H 139D 6N 5O 5 Chemical formula: C 70 H 139 D 6 N 5 O 5
分子量：1143.01Molecular weight: 1143.01
化合物71可根据实施例1描述的代表性路线合成。Compound 71 can be synthesized according to the representative route described in Example 1.
H NMR(300MHz,DMSO-d6):1H NMR(DMSO-d6):δ(ppm)5.4(s,5H),3.49(m,5H,),2.22-2.71(m,22H),1.47(m,10H),1.28(m,80H),0.90(m,10H).H NMR(300MHz,DMSO-d6):1H NMR(DMSO-d6):δ(ppm)5.4(s,5H),3.49(m,5H,),2.22-2.71(m,22H),1.47(m, 10H),1.28(m,80H),0.90(m,10H).
实施例73：Example 73:
化合物72的合成Synthesis of Compound 72
Figure PCTCN2022084347-appb-000089
Figure PCTCN2022084347-appb-000089
化学式：C 70H 139D 6N 5O 5 Chemical formula: C 70 H 139 D 6 N 5 O 5
分子量：1143.01Molecular weight: 1143.01
化合物72可根据实施例1描述的代表性路线合成。Compound 72 can be synthesized according to the representative route described in Example 1.
H NMR(300MHz,DMSO-d6):1H NMR(DMSO-d6):δ(ppm)5.4(s,5H),3.49(m,5H,),2.22-2.71(m,22H),1.47(m,10H),1.28(m,80H),0.90(m,10H).H NMR(300MHz,DMSO-d6):1H NMR(DMSO-d6):δ(ppm)5.4(s,5H),3.49(m,5H,),2.22-2.71(m,22H),1.47(m, 10H),1.28(m,80H),0.90(m,10H).
实施例74：Example 74:
化合物73的合成Synthesis of Compound 73
Figure PCTCN2022084347-appb-000090
Figure PCTCN2022084347-appb-000090
化学式：C 70H 139D 6N 5O 5 Chemical formula: C 70 H 139 D 6 N 5 O 5
分子量：1143.01Molecular weight: 1143.01
化合物73可根据实施例1描述的代表性路线合成。Compound 73 can be synthesized according to the representative route described in Example 1.
H NMR(300MHz,DMSO-d6):1H NMR(DMSO-d6):δ(ppm)5.4(s,5H),3.49(m,5H,),2.22-2.71(m,22H),1.47(m,10H),1.28(m,80H),0.90(m,10H).H NMR(300MHz,DMSO-d6):1H NMR(DMSO-d6):δ(ppm)5.4(s,5H),3.49(m,5H,),2.22-2.71(m,22H),1.47(m, 10H),1.28(m,80H),0.90(m,10H).
实施例75：Example 75:
化合物74的合成Synthesis of Compound 74
Figure PCTCN2022084347-appb-000091
Figure PCTCN2022084347-appb-000091
化学式：C 70H 139D 6N 5O 5 Chemical formula: C 70 H 139 D 6 N 5 O 5
分子量：1143.01Molecular weight: 1143.01
化合物74可根据实施例1描述的代表性路线合成。Compound 74 can be synthesized according to the representative route described in Example 1.
H NMR(300MHz,DMSO-d6):1H NMR(DMSO-d6):δ(ppm)5.4(s,5H),3.49(m,5H,),2.22-2.71(m,22H),1.47(m,10H),1.28(m,80H),0.90(m,10H).H NMR(300MHz,DMSO-d6):1H NMR(DMSO-d6):δ(ppm)5.4(s,5H),3.49(m,5H,),2.22-2.71(m,22H),1.47(m, 10H),1.28(m,80H),0.90(m,10H).
实施例76：Example 76:
化合物75的合成Synthesis of compound 75
Figure PCTCN2022084347-appb-000092
Figure PCTCN2022084347-appb-000092
化学式：C 70H 138D 7N 5O 5 Chemical formula: C 70 H 138 D 7 N 5 O 5
分子量：1144.01Molecular weight: 1144.01
化合物75可根据实施例1描述的代表性路线合成。Compound 75 can be synthesized according to the representative route described in Example 1.
H NMR(300MHz,DMSO-d6):1H NMR(DMSO-d6):δ(ppm)5.4(s,5H),3.49(m,5H,),2.22-2.71(m,21H),1.47(m,10H),1.28(m,80H),0.90(m,10H).H NMR(300MHz,DMSO-d6):1H NMR(DMSO-d6):δ(ppm)5.4(s,5H),3.49(m,5H,),2.22-2.71(m,21H),1.47(m, 10H),1.28(m,80H),0.90(m,10H).
实施例77：Example 77:
化合物76的合成Synthesis of compound 76
Figure PCTCN2022084347-appb-000093
Figure PCTCN2022084347-appb-000093
化学式：C 70H 138D 7N 5O 5 Chemical formula: C 70 H 138 D 7 N 5 O 5
分子量：1144.01Molecular weight: 1144.01
化合物76可根据实施例1描述的代表性路线合成。Compound 76 can be synthesized according to the representative route described in Example 1.
H NMR(300MHz,DMSO-d6):1H NMR(DMSO-d6):δ(ppm)5.4(s,5H),3.49(m,5H,),2.22-2.71(m,21H),1.47(m,10H),1.28(m,80H),0.90(m,10H).H NMR(300MHz,DMSO-d6):1H NMR(DMSO-d6):δ(ppm)5.4(s,5H),3.49(m,5H,),2.22-2.71(m,21H),1.47(m, 10H),1.28(m,80H),0.90(m,10H).
实施例78：Example 78:
化合物77的合成Synthesis of compound 77
Figure PCTCN2022084347-appb-000094
Figure PCTCN2022084347-appb-000094
化学式：C 70H 138D 7N 5O 5 Chemical formula: C 70 H 138 D 7 N 5 O 5
分子量：1144.01Molecular weight: 1144.01
化合物77可根据实施例1描述的代表性路线合成。Compound 77 can be synthesized according to the representative route described in Example 1.
H NMR(300MHz,DMSO-d6):1H NMR(DMSO-d6):δ(ppm)5.4(s,5H),3.49(m,5H,),2.22-2.71(m,21H),1.47(m,10H),1.28(m,80H),0.90(m,10H).H NMR(300MHz,DMSO-d6):1H NMR(DMSO-d6):δ(ppm)5.4(s,5H),3.49(m,5H,),2.22-2.71(m,21H),1.47(m, 10H),1.28(m,80H),0.90(m,10H).
实施例79：Example 79:
化合物78的合成Synthesis of Compound 78
Figure PCTCN2022084347-appb-000095
Figure PCTCN2022084347-appb-000095
化学式：C 70H 138D 7N 5O 5 Chemical formula: C 70 H 138 D 7 N 5 O 5
分子量：1144.01Molecular weight: 1144.01
化合物78可根据实施例1描述的代表性路线合成。Compound 78 can be synthesized according to the representative route described in Example 1.
H NMR(300MHz,DMSO-d6):1H NMR(DMSO-d6):δ(ppm)5.4(s,5H),3.49(m,5H,),2.22-2.71(m,21H),1.47(m,10H),1.28(m,80H),0.90(m,10H).H NMR(300MHz,DMSO-d6):1H NMR(DMSO-d6):δ(ppm)5.4(s,5H),3.49(m,5H,),2.22-2.71(m,21H),1.47(m, 10H),1.28(m,80H),0.90(m,10H).
实施例80Example 80
利用脂质纳米颗粒组合物的荧光素酶mRNA体内评价In vivo evaluation of luciferase mRNA using a lipid nanoparticle composition
将阳离子脂质、DSPC、胆固醇和PEG-脂质以50:10:38:2或48:10:40:2的摩尔比溶解在乙醇中。以约10:1至30:1的总脂质与mRNA的重量比制备脂质纳米颗粒(LNP)。简而言之，将mRNA在10ml至50ml柠檬酸盐缓冲液(pH＝4)稀释至0.15mg/mL。使用注射器泵，将脂质的乙醇溶液与mRNA水溶液以约1:5至1:3(体积/体积)的比例混合，总流速为10ml/min以上。然后去除乙醇，并通过透析用PBS替代外部的缓冲液。最后，将脂质纳米颗粒通过0.2um孔径的无菌过滤器过滤。使用Malvern Zetasizer Nano ZS通过准弹性光散射测定的脂质纳米颗粒的粒径为直径大约65-105nm，并且在一些情况下，直径大约75-100nm。Cationic lipids, DSPC, cholesterol and PEG-lipids were dissolved in ethanol at a molar ratio of 50:10:38:2 or 48:10:40:2. Lipid nanoparticles (LNPs) were prepared at a weight ratio of total lipid to mRNA of about 10:1 to 30:1. Briefly, mRNA was diluted to 0.15 mg/mL in 10 ml to 50 ml citrate buffer (pH=4). Using a syringe pump, mix the ethanolic lipid solution with the aqueous mRNA solution at a ratio of about 1:5 to 1:3 (vol/vol) at a total flow rate of 10 ml/min or more. Ethanol was then removed and the external buffer was replaced by PBS by dialysis. Finally, the lipid nanoparticles were filtered through a sterile filter with a pore size of 0.2 μm. The particle size of the lipid nanoparticles, as determined by quasi-elastic light scattering using a Malvern Zetasizer Nano ZS, is approximately 65-105 nm in diameter, and in some cases, approximately 75-100 nm in diameter.
根据国家科学技术委员会制定的指南，在6-8周龄的雌性C57BL/6小鼠，8-10周龄CD-1小鼠上进行研究。通过尾静脉注射全身性给予不同剂量的mRNA脂质纳米颗粒，并在给药后的特定时间点(例如5小时)使动物安乐死。将肝脏和脾脏收集在预先称重的管中，确定重量，立即在液氮中快速冷冻，并且在-80℃下储存，直至用于分析。The study was performed on 6-8 week old female C57BL/6 mice, 8-10 week old CD-1 mice according to the guidelines established by the National Science and Technology Commission. Different doses of mRNA lipid nanoparticles were administered systemically via tail vein injection, and animals were euthanized at specific time points (eg, 5 hours) after administration. Livers and spleens were collected in pre-weighed tubes, weight determined, immediately snap frozen in liquid nitrogen, and stored at -80°C until analysis.
对于肝脏，切割约50mg以便在2mL FastPrep管(MP Biomedicals，Solon OH)中进行分析。向各个管中加入1/4"陶瓷球(MP Biomedicals)，并将平衡至室温的500μL的Glo裂解缓冲液-GLB(Promega，Madison WI)加入到肝脏组织中。使用FastPrep24仪器(MP Biomedicals)将肝脏组织在2x6.0m/s下均匀化15秒。将匀浆在室温下孵育5分钟，然后在GLB中进行1:4稀释，并使用SteadyGlo荧光素酶测定系统(Promega)进行评估。具体地，将50uL的稀释的组织匀浆与50μL的SteadyGlo底物反应，摇振10秒，接着孵育5分钟，然后使用SpectraMAX_L化学发光型酶标仪(美谷分子仪器(上海)有限公司)定量。通过使用BCA蛋白质定量试剂盒(上海易色医疗科技有限公司)来确定测定的蛋白质的量。然后将相对发光度单位(RLU)归一化成所测定蛋白质的总ug。为了将RLU转化成μg荧光素酶，用QuantiLum重组荧光素酶(Promega)生成了标准曲线。For liver, approximately 50 mg was cut for analysis in 2 mL FastPrep tubes (MP Biomedicals, Solon OH). 1/4" ceramic balls (MP Biomedicals) were added to each tube, and 500 μL of Glo Lysis Buffer-GLB (Promega, Madison WI) equilibrated to room temperature was added to the liver tissue. Using the FastPrep24 instrument (MP Biomedicals), the Liver tissue was homogenized at 2x6.0m/s for 15 s. The homogenate was incubated at room temperature for 5 min, then diluted 1:4 in GLB and evaluated using the SteadyGlo luciferase assay system (Promega). Specifically , 50uL of diluted tissue homogenate was reacted with 50μL of SteadyGlo substrate, shaken for 10 seconds, then incubated for 5 minutes, and then quantified using a SpectraMAX_L chemiluminescent microplate reader (Megu Molecular Instruments (Shanghai) Co., Ltd.). Use the BCA Protein Quantification Kit (Shanghai Yise Medical Technology Co., Ltd.) to determine the amount of assayed protein. The relative luminescence units (RLU) are then normalized to the total ug of assayed protein. In order to convert RLU into μg fluorescein Enzymes, standard curves were generated using QuantiLum recombinant luciferase (Promega).
来自Trilink Biotechnologies的FLuc mRNA(L-6107)将表达荧光素酶蛋白，其最初从萤火虫(Photinus pyralis)中分离出来。Fluc通常用于哺乳动物细胞培养物中以测量基因表达和细胞活力。其在底物萤光素存在下发射出生物性光。这种加帽并且聚腺昔酸化的mRNA被5-甲基胞苷和假尿苷完全取代。FLuc mRNA (L-6107) from Trilink Biotechnologies will express the luciferase protein, which was originally isolated from the firefly (Photinus pyralis). Fluc is commonly used in mammalian cell culture to measure gene expression and cell viability. It emits bioluminescence in the presence of the substrate luciferin. This capped and polyadenylated mRNA is completely replaced by 5-methylcytidine and pseudouridine.
实施例65Example 65
所配制得脂质的pKa的测定Determination of the pKa of the prepared lipid
所配制的阳离子脂质的pKa与用于递送核酸的LNP的效果相关。优选的pKa范围是5～7。使用基于2-(对甲苯胺基)-6-荼磺酸(TNS)的荧光的分析，在脂质纳米颗粒中测定各阳离子脂质的pKa。如实施例64中所述，使用有序的方法来制备在PBS中的浓度为0.4mM总脂质的包含阳离子脂质/DSPC/胆固醇/PEG 脂质(50/10/38/2mol％)的脂质纳米颗粒。将TNS在蒸馏水中制备成100uM储备溶液。将囊泡稀释成在2mL缓冲溶液中含24uM脂质，所述缓冲溶液含有10mM HEPES、10mM MES、10mM乙酸按、130mM NaCl，其中PH值为2.5～11。加入等份的TNS溶液以产生l uM的终浓度，并且在涡旋混合之后，在室温下使用321nm和445nm的激发波长和发射波长在SLM Aminco Series 2发光分光光度计中测量荧光强度。对荧光数据应用S形最佳拟合分析，并将pKa测量为产生半数最大荧光强度的pH。The pKa of the formulated cationic lipid correlates with the effectiveness of the LNP for delivery of nucleic acids. The preferred pKa range is 5-7. The pKa of each cationic lipid was determined in lipid nanoparticles using an assay based on the fluorescence of 2-(p-toluidino)-6-phthanesulfonic acid (TNS). As described in Example 64, a sequenced method was used to prepare cationic lipids/DSPC/cholesterol/PEG lipids (50/10/38/2 mol%) at a concentration of 0.4 mM total lipids in PBS. Lipid nanoparticles. TNS was prepared as a 100 uM stock solution in distilled water. The vesicles were diluted to contain 24uM lipid in 2mL buffer solution containing 10mM HEPES, 10mM MES, 10mM sodium acetate, 130mM NaCl, wherein the pH value was 2.5-11. Aliquots of TNS solution were added to give a final concentration of 1 uM, and after vortex mixing, fluorescence intensity was measured in a SLM Aminco Series 2 Luminescence Spectrophotometer at room temperature using excitation and emission wavelengths of 321 nm and 445 nm. Sigmoid best-fit analysis was applied to the fluorescence data, and pKa was measured as the pH that yielded half the maximum fluorescence intensity.
实施例66Example 66
使用体内荧光素酶mRNA表达的啮齿动物模型测定含有各种阳离子脂质的脂质纳米颗粒制剂的效能Determination of the Potency of Lipid Nanoparticle Formulations Containing Various Cationic Lipids Using a Rodent Model of In Vivo Luciferase mRNA Expression
为了比较的目的，如实施例64所述，使用有序混合方法，将这些脂质也用于配制含有FLuc mRNA(L-6107)的脂质纳米颗粒。使用以下摩尔比来配制脂质纳米颗粒:50％阳离子脂质/10％二硬脂酰磷脂酰胆碱(DSPC)/38％胆固醇/2％PEG脂质("PEG-DMG"，即，(1-(单甲氧基一聚乙二醇)-2，3一二肉豆蔻酰基甘油，平均PEG分子量为2000)。如实施例64所述，在经由尾静脉注射施用之后的5小时，通过测量肝脏中的荧光素酶表达来确定相对活性。在0.3和1.0mg mRNA/kg的剂量下比较所述活性，并表达成在如实施例64所述的施用之后5小时测量的ng荧光素酶/g肝脏。实施例65及66结果如表2所示。For comparison purposes, these lipids were also used to formulate lipid nanoparticles containing FLuc mRNA (L-6107) using the ordered mixing method as described in Example 64. Lipid nanoparticles were formulated using the following molar ratios: 50% cationic lipid/10% distearoylphosphatidylcholine (DSPC)/38% cholesterol/2% PEG lipid ("PEG-DMG", i.e., ( 1-(monomethoxy-polyethylene glycol)-2,3-dimyristoylglycerol, the average PEG molecular weight is 2000). As described in Example 64, 5 hours after administration via tail vein injection, by Relative activity was determined by measuring luciferase expression in the liver. The activity was compared at doses of 0.3 and 1.0 mg mRNA/kg and expressed as ng luciferase measured 5 hours after administration as described in Example 64 /g liver.The results of embodiment 65 and 66 are shown in table 2.
表2与mRNA表现出活性的比较脂质Table 2. Comparative lipids exhibiting activity with mRNA
Figure PCTCN2022084347-appb-000096
Figure PCTCN2022084347-appb-000096
Figure PCTCN2022084347-appb-000097
Figure PCTCN2022084347-appb-000097
Figure PCTCN2022084347-appb-000098
Figure PCTCN2022084347-appb-000098
Figure PCTCN2022084347-appb-000099
Figure PCTCN2022084347-appb-000099
Figure PCTCN2022084347-appb-000100
Figure PCTCN2022084347-appb-000100
Figure PCTCN2022084347-appb-000101
Figure PCTCN2022084347-appb-000101
Figure PCTCN2022084347-appb-000102
Figure PCTCN2022084347-appb-000102
Figure PCTCN2022084347-appb-000103
Figure PCTCN2022084347-appb-000103
以上所述实施例的各技术特征可以进行任意的组合，为使描述简洁，未对上述实施例中的各个技术特征所有可能的组合都进行描述，然而，只要这些技术特征的组合不存在矛盾，都应当认为是本说明书记载的范围。The technical features of the above-mentioned embodiments can be combined arbitrarily. To make the description concise, all possible combinations of the technical features in the above-mentioned embodiments are not described. However, as long as there is no contradiction in the combination of these technical features, should be considered as within the scope of this specification.
以上所述实施例仅表达了本发明的几种实施方式，其描述较为具体和详细，但并不能因此而理解为对公开专利范围的限制。应当指出的是，对于本领域的普通技术人员来说，在不脱离本发明构思的前提下，还可以做出若干变形和改进，这些都属于本发明的保护范围。因此，本发明专利的保护范围应以所附权利要求为准。The above-mentioned embodiments only express several implementation modes of the present invention, and the descriptions thereof are relatively specific and detailed, but should not be construed as limiting the scope of the disclosed patents. It should be noted that those skilled in the art can make several modifications and improvements without departing from the concept of the present invention, and these all belong to the protection scope of the present invention. Therefore, the protection scope of the patent for the present invention should be based on the appended claims.

Claims (15)

  1. 一种式“I”化合物:A compound of formula "I":
    Figure PCTCN2022084347-appb-100001
    Figure PCTCN2022084347-appb-100001
    或其N-氧化物，或其盐或异构体。or its N-oxide, or its salt or isomer.
    其中结构式“I”中的R 1、R 2、R 3、和R 4均为独立的2个“氢”同位素(包括同位素“氕”和“氘”)的组合，具体表现为“HH”、“HD”和“DD”的组合； Among them, R 1 , R 2 , R 3 , and R 4 in the structural formula "I" are all independent combinations of two "hydrogen" isotopes (including the isotopes "protium" and "deuterium"), specifically expressed as "HH", A combination of "HD" and "DD";
    R 1和R 2独立地选自由“HH”、“HD”和“DD”组成的任意一组； R and R are independently selected from any group consisting of "HH", "HD" and "DD";
    R 3和R 4独立地选自由“HH、HD”、“HD、HH”、“HH、DD”“DD、HH”“HD、DD”和“DD、DD”组成的任意一组； R3 and R4 are independently selected from any group consisting of "HH, HD", "HD, HH", "HH, DD", "DD, HH", "HD, DD" and "DD, DD";
    R 1、R 2、R 3、和R 4均有独立性，但不能同时为“HH”，即至少含有一个“HD”的组合，具体组合情况分8种情况，包括含1个“D”的组合、含2个“D”的组合、含3个“D”的组合、含4个“D”的组合、含5个“D”的组合、含6个“D”的组合、含7个“D”的组合以及含8个“D”的组合； R 1 , R 2 , R 3 , and R 4 are all independent, but they cannot be "HH" at the same time, that is, a combination containing at least one "HD". There are 8 specific combinations, including one "D" combination of 2 "D", combination of 3 "D", combination of 4 "D", combination of 5 "D", combination of 6 "D", combination of 7 A combination of "D"s and a combination of 8 "Ds";
  2. 选自表1中的化合物的化合物。A compound selected from the compounds in Table 1.
  3. 包含权利要求1～2中任一项所述的化合物和治疗剂和/或预防剂的组合物。A composition comprising a compound according to any one of claims 1-2 and a therapeutic agent and/or a preventive agent.
  4. 如权利要求3所述的组合物，其还包含选自中性脂质、类固醇以及聚合 物缀合的脂质中的一种或多种赋形剂。The composition of claim 3, further comprising one or more excipients selected from neutral lipids, steroids, and polymer-conjugated lipids.
  5. 如前述权利要求4所述的组合物，其中所述组分中的中性脂质选自以下的一种或多种物质混合：1，2-二硬脂酰基-sn-甘油-3-磷酸胆碱(DSPC)、1，2-二棕榈酰基-sn-甘油-3-磷酸胆碱(DPPC)、1，2-二肉豆寇基-sn-甘油-磷酸胆碱(DMPC)、1，2-二油酰基-sn-甘油-3-磷酸胆碱(DOPC)、1-棕榈酰基-2-油酰基-sn-甘油-3-磷酸胆碱(POPC)、1，2-二油酰基-sn-甘油-3-磷酸乙醇胺(DOPE)和鞘磷脂(SM)。The composition as claimed in claim 4, wherein the neutral lipid in the component is selected from one or more mixtures of the following: 1,2-distearoyl-sn-glycero-3-phosphate Choline (DSPC), 1,2-Dipalmitoyl-sn-glycero-3-phosphocholine (DPPC), 1,2-Dimyristyl-sn-glycerol-phosphocholine (DMPC), 1, 2-Dioleoyl-sn-glycero-3-phosphocholine (DOPC), 1-palmitoyl-2-oleoyl-sn-glycero-3-phosphocholine (POPC), 1,2-dioleoyl- sn-glycero-3-phosphoethanolamine (DOPE) and sphingomyelin (SM).
  6. 如前述权利要求5所述的组合物，其中所述中性脂质为DSPC。The composition of claim 5, wherein the neutral lipid is DSPC.
  7. 如前述权利要求4～6所述的组合物，其中所述组分中的类固醇选自以下一种或多种物质混合：胆固醇、粪固醇、谷固醇、麦角固醇、菜油固醇、豆固醇、菜籽固醇、番茄碱、熊果酸、α-生育酚。The composition as claimed in claims 4-6, wherein the steroid in the component is selected from one or more of the following mixtures: cholesterol, fecal sterol, sitosterol, ergosterol, campesterol, Stigmasterol, brassicasterol, tomatine, ursolic acid, alpha-tocopherol.
  8. 如前述权利要求7所述的组合物，其中所述类固醇为胆固醇。A composition as claimed in claim 7, wherein said steroid is cholesterol.
  9. 如前述权利要求4～8所述的组合物，其中所述组分中的聚合物缀合的脂质为聚乙二醇化脂质。The composition according to the preceding claims 4-8, wherein the polymer-conjugated lipid in said component is a pegylated lipid.
  10. 如前述权利要求中9所述的组合物，其中所述聚乙二醇化脂质为1,2-二肉豆蔻酰基-sn-甘油甲氧基聚乙二醇(PEG-DMG)A composition as claimed in claim 9, wherein the pegylated lipid is 1,2-dimyristoyl-sn-glycerol methoxypolyethylene glycol (PEG-DMG)
  11. 如前述权利要求中任一项所述的组合物，其中所述治疗剂和/或预防剂是能够引起免疫响应的疫苗或化合物，其中包括核酸。The composition of any one of the preceding claims, wherein the therapeutic and/or prophylactic agent is a vaccine or compound capable of eliciting an immune response, including nucleic acids.
  12. 如前述权利要求11的组合物，其中所述核酸为RNA，其选自以下组成：siRNA、aiRNA、miRNA、dsRNA、shRNA、mRNA以及其混合物。The composition of claim 11, wherein the nucleic acid is RNA selected from the group consisting of siRNA, aiRNA, miRNA, dsRNA, shRNA, mRNA and mixtures thereof.
  13. 如前述权利要求12的组合物，其中所述RNA为mRNA。The composition of claim 12, wherein said RNA is mRNA.
  14. 向有需要的受试者施用治疗剂和/或预防剂的方法，所述方法包括制备或提供上述权利要求中任一项所述的组合物，并向所述对象施用所述组合物。A method of administering a therapeutic and/or prophylactic agent to a subject in need thereof, the method comprising preparing or providing a composition according to any one of the preceding claims, and administering the composition to the subject.
  15. 如前述权利要求中任一项所述的受试者为哺乳动物或人。The subject of any one of the preceding claims is a mammal or a human.
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