WO2024022507A1 - 一种包含kras g12d抑制剂的药物组合物 - Google Patents

一种包含kras g12d抑制剂的药物组合物 Download PDF

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Publication number
WO2024022507A1
WO2024022507A1 PCT/CN2023/109901 CN2023109901W WO2024022507A1 WO 2024022507 A1 WO2024022507 A1 WO 2024022507A1 CN 2023109901 W CN2023109901 W CN 2023109901W WO 2024022507 A1 WO2024022507 A1 WO 2024022507A1
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Prior art keywords
cancer
pharmaceutical composition
composition according
liposome
ammonium
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PCT/CN2023/109901
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English (en)
French (fr)
Inventor
沙瑞霖
段子卿
王海凤
仝新勇
王萍萍
冯标
王斌
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Shanghai Hengrui Pharmaceutical Co Ltd
Jiangsu Hengrui Pharmaceutical Co Ltd
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Shanghai Hengrui Pharmaceutical Co Ltd
Jiangsu Hengrui Pharmaceutical Co Ltd
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Priority to AU2023314335A priority Critical patent/AU2023314335A1/en
Priority to CA3261035A priority patent/CA3261035A1/en
Priority to KR1020257005566A priority patent/KR20250037564A/ko
Priority to EP23845711.3A priority patent/EP4563151A4/en
Priority to CN202380046186.1A priority patent/CN119384279A/zh
Priority to JP2025501568A priority patent/JP2025524799A/ja
Publication of WO2024022507A1 publication Critical patent/WO2024022507A1/zh
Priority to MX2025001036A priority patent/MX2025001036A/es
Anticipated expiration legal-status Critical
Ceased legal-status Critical Current

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    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P35/00—Antineoplastic agents
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K31/00—Medicinal preparations containing organic active ingredients
    • A61K31/33—Heterocyclic compounds
    • A61K31/395—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins
    • A61K31/55—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having seven-membered rings, e.g. azelastine, pentylenetetrazole
    • A61K31/553—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having seven-membered rings, e.g. azelastine, pentylenetetrazole having at least one nitrogen and one oxygen as ring hetero atoms, e.g. loxapine, staurosporine
    • 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
    • A61K9/00—Medicinal preparations characterised by special physical form
    • A61K9/0012—Galenical forms characterised by the site of application
    • A61K9/0019—Injectable compositions; Intramuscular, intravenous, arterial, subcutaneous administration; Compositions to be administered through the skin in an invasive manner
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K9/00—Medicinal preparations characterised by special physical form
    • A61K9/10—Dispersions; Emulsions
    • A61K9/127—Synthetic bilayered vehicles, e.g. liposomes or liposomes with cholesterol as the only non-phosphatidyl surfactant
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K9/00—Medicinal preparations characterised by special physical form
    • A61K9/10—Dispersions; Emulsions
    • A61K9/127—Synthetic bilayered vehicles, e.g. liposomes or liposomes with cholesterol as the only non-phosphatidyl surfactant
    • A61K9/1271—Non-conventional liposomes, e.g. PEGylated liposomes or liposomes coated or grafted with polymers
    • 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
    • A61K9/14—Particulate form, e.g. powders, Processes for size reducing of pure drugs or the resulting products, Pure drug nanoparticles
    • A61K9/19—Particulate form, e.g. powders, Processes for size reducing of pure drugs or the resulting products, Pure drug nanoparticles lyophilised, i.e. freeze-dried, solutions or dispersions
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P35/00—Antineoplastic agents
    • A61P35/02—Antineoplastic agents specific for leukemia

Definitions

  • the present disclosure belongs to the field of pharmaceutical preparations, and specifically relates to a pharmaceutical composition containing a KRAS G12D inhibitor and a preparation method thereof.
  • RAS is one of the oncogenes with the highest mutation rate in tumors. About 30% of human malignant tumors are related to mutations in the RAS gene.
  • the RAS family includes KRAS, NRAS and HRAS, of which KRAS mutations are the most common, accounting for approximately 85%.
  • KRAS mutations are common in solid tumors, with high frequency mutations found in the three most lethal cancers in humans: lung cancer (17%), colorectal cancer (33%), and pancreatic cancer (61%).
  • 97% are mutations in the 12th or 13th amino acid residue, among which G12D is an important mutation.
  • Data analysis of European and American populations shows that in pancreatic cancer, colorectal cancer and non-small cell lung cancer, G12D mutations account for 36%, 12% and 4% of patients respectively.
  • KRAS After KRAS is activated, it regulates cell proliferation, survival, migration, metabolism and other functions through numerous downstream signaling pathways represented by RAF-MEK-ERK, PI3K-AKT-mTOR and TIAM1-RAc. After the KRAS gene is mutated, the protein remains in an activated state, resulting in continued activation of downstream signaling pathways and promoting tumorigenesis.
  • KRAS protein lacks small molecule binding sites in the traditional sense and has ultra-high affinity to guanylate, making it extremely difficult to inhibit, it has long been considered an undruggable drug target.
  • KRAS has been and remains a target of great concern for drug development.
  • G12C inhibitors there is still a lack of KRAS inhibitors that are effective against other mutations, leaving most patients with KRAS mutations still untreatable.
  • G12D as a mutant that is widely expressed in a variety of tumors, the development of inhibitors against it has important clinical significance.
  • WO2022268051 provides a new KRAS G12D inhibitor compound (Formula I), which has good pharmaceutical activity.
  • It is an object of the present disclosure to provide a pharmaceutical composition comprising a compound of formula I or a pharmaceutically acceptable salt thereof.
  • One aspect of the present disclosure provides a pharmaceutical composition
  • a pharmaceutical composition comprising a compound of Formula I or a pharmaceutically acceptable salt thereof and a lipid
  • the lipid includes at least one phospholipid.
  • compositions of the present disclosure are liposomes.
  • Another aspect of the present disclosure provides a liposome comprising a compound of Formula I or a pharmaceutically acceptable salt thereof.
  • Lipids of the present disclosure may include completely neutral or negatively charged phospholipids.
  • phospholipid refers to a hydrophobic molecule containing at least one phosphorus group, which may be natural or synthetic.
  • phospholipids may contain phosphorus-containing groups and saturated or unsaturated alkyl groups optionally substituted with OH, COOH, oxo, amine, or substituted or unsubstituted aryl groups.
  • Phospholipids differ from each other in the length and degree of unsaturation of their acyclic (acylic) chains.
  • the phospholipid includes one or more of phosphatidylcholine, phosphatidylethanolamine, phosphatidylglycerol, phosphatidylserine, phosphatidic acid, and phosphatidylinositol.
  • phosphatidylcholine refers to phosphatidylcholine and its derivatives.
  • phospholipids suitable for use in the present disclosure include dipalmitoylphosphatidylcholine (DPPC), distearoylphosphatidylcholine (DSPC), dimyristoylphosphatidylcholine (DMPC), 1-palmitoyl- 2-linoleoyl-sn-glycerol-3-phosphatidylcholine (PLPC), dioleoylphosphatidylcholine (DOPC), dierucoylphosphatidylcholine (DEPC), egg yolk phosphatidylcholine (EPC) , dilauroylphosphatidylcholine (DLPC), hydrogenated soybean phosphatidylcholine (HSPC), l-myristoyl-2-palmitoylphosphatidylcholine (MPPC), l-palmitoyl-2-myristoyl Phosphatidylcholine (PMPC), l-palmitoyl-2-stearoylphosphatidylcholine (
  • the hydrophilic polymer-modified phospholipids include, but are not limited to, polyethylene glycol-modified distearoylphosphatidylethanolamine (PEG-DSPE), polyethylene glycol-modified distearoylphosphatidylglycerol (PEG-DSPG). ), polyethylene glycol modified cholesterol (PEG-CHOL).
  • PEG-DSPE polyethylene glycol-modified distearoylphosphatidylethanolamine
  • PEG-DSPG polyethylene glycol-modified distearoylphosphatidylglycerol
  • PEG-CHOL polyethylene glycol modified cholesterol
  • the phospholipids described in this disclosure are selected from the group consisting of dipalmitoylphosphatidylcholine (DPPC), distearoylphosphatidylcholine (DSPC), dioleoylphosphatidylcholine (DOPC), Erucylphosphatidylcholine (DEPC), dilauroylphosphatidylcholine (DLPC), hydrogenated soybean phosphatidylcholine (HSPC), distearoylphosphatidylethanolamine (DSPE) and dimyristoylphosphatidylcholine (DMPC) and their polyethylene glycol-modified derivatives.
  • DPPC dipalmitoylphosphatidylcholine
  • DSPC distearoylphosphatidylcholine
  • DOPC dioleoylphosphatidylcholine
  • DEPC Erucylphosphatidylcholine
  • DLPC dilauroylphosphatidylcholine
  • HSPC hydrogenated soybean phosphat
  • the liposomes contain polyethylene glycol modified distearoylphosphatidylethanolamine.
  • the molecular weight of polyethylene glycol in the polyethylene glycol-modified distearoylphosphatidylethanolamine ranges from 200 to 10,000 daltons. In some embodiments, the molecular weight of polyethylene glycol in the polyethylene glycol-modified distearoylphosphatidylethanolamine is 1000-5000 daltons. In some embodiments, the molecular weight of polyethylene glycol in the polyethylene glycol-modified distearoylphosphatidylethanolamine is 2000-5000 daltons.
  • the polyethylene glycol modified distearoylphosphatidylethanolamine is selected from polyethylene glycol 2000-distearoylphosphatidylethanolamine (mPEG2000-DSPE).
  • the phospholipid molar content is selected from 10 to 80%, including but not limited to 10%, 11%, 12%, 13%, 14%, 15%, 16%, relative to the amount of the total lipid mixture. %, 17%, 18%, 19%, 20%, 21%, 22%, 23%, 24%, 25%, 26%, 27%, 28%, 29%, 30%, 31%, 32%, 33%, 34%, 35%, 36%, 37%, 38%, 39%, 40%, 41%, 42%, 43%, 44%, 45%, 46%, 47%, 48%, 49% ,50%,51%,52%,53%,54%,55%,56%,57%,58%,59%,60%,61%,62%,63%,64%,65%,66 %, 67%, 68%, 69%, 70%, 71%, 72%, 73%, 74%, 75%, 76%, 77%, 78%, 79%, 80% or any value between any two values ,
  • the present disclosure may also include other neutral lipids, cationic lipids, and/or anionic lipids.
  • Examples of other neutral lipids useful in the present disclosure include: one or more of steroids such as cholesterol and its derivatives, lecithin, soybean lecithin, cephalin, sphingomyelin, hydrogenated soybean lecithin.
  • steroids such as cholesterol and its derivatives, lecithin, soybean lecithin, cephalin, sphingomyelin, hydrogenated soybean lecithin.
  • the molar percentage of steroid relative to the amount of the total lipid mixture does not exceed 90%.
  • the lipid includes at least one phospholipid and cholesterol.
  • the molar percentage of cholesterol relative to the amount of the total lipid mixture may be from 0.1% to 90%, for example from 10% to 80%, or from 20% to 70%.
  • the phospholipid is selected from dipalmitoylphosphatidylcholine (DPPC), distearoylphosphatidylcholine (DSPC), dioleoylphosphatidylcholine (DOPC), dierucylphospholipid Acylcholine (DEPC), di Lauroylphosphatidylcholine (DLPC), hydrogenated soybean phosphatidylcholine (HSPC), distearoylphosphatidylethanolamine (DSPE) and dimyristoylphosphatidylcholine (DMPC) and their polyethylene glycol modifications Derivatives.
  • DPPC dipalmitoylphosphatidylcholine
  • DSPC distearoylphosphatidylcholine
  • DOPC dioleoylphosphatidylcholine
  • DEPC dierucylphospholipid Acylcholine
  • DLPC di Lauroylphosphatidylcholine
  • HSPC hydrogenated soybean phosphatidylcholine
  • DSPE distea
  • the lipids comprise hydrogenated soy phosphatidylcholine, cholesterol, and polyethylene glycol 2000-distearoylphosphatidylethanolamine.
  • the hydrophilic polymer-modified phospholipid derivative content (molar fraction relative to the total liposome membrane components) is selected from 0.1 to 50%, including but not limited to 0.1%, 0.2%, 0.3% ,0.4%,0.5%,0.6%,0.7%,0.8%,0.9%,1%,2%,3%,4%,5%,6%,7%,8%,9%,10%,11 %, 12%, 13%, 14%, 15%, 16%, 17%, 18%, 19%, 20%, 21%, 22%, 23%, 24%, 25%, 26%, 27%, 28%, 29%, 30%, 31%, 32%, 33%, 34%, 35%, 36%, 37%, 38%, 39%, 40%, 41%, 42%, 43%, 44% , 45%, 46%, 47%, 48%, 49%, 50% or any value between any two values, for example, it can be 0.5 to 20%.
  • the pharmaceutical composition further comprises a capture agent.
  • the aqueous phase within the liposome contains a capture agent.
  • the preparation method of the liposome can adopt an active drug loading method.
  • the active drug loading method is a drug loading method that transports therapeutic agents from the external medium across the double-layer membrane of the liposome to the internal aqueous space through a polyatomic ion-gradient. These gradients work by embedding at least one polyatomic ion as a capture agent in the internal aqueous space of the liposomes, and using an additional medium with a lower polyatomic ion concentration, such as pure water, sucrose solution or physiological saline. Produced by displacing the external medium of liposomes through known techniques such as column separation, dialysis or centrifugation. A polyatomic ion gradient is created between the internal aqueous space of the liposome and the external medium to trap the therapeutic agent in the internal aqueous space of the liposome.
  • Capture agents described in the present disclosure include, but are not limited to, sulfates, sulfites, gluconates, phosphates, hydrogen phosphates, molybdates, carbonates, nitrates, and the like.
  • Exemplary capture agents include, but are not limited to, ammonium sulfate, ammonium phosphate, ammonium molybdate, ammonium sucrose octasulfate, triethylammonium sucrose octasulfate, copper gluconate, ammonium ethyl sulfate, ammonium ethylenediaminetetraacetate, Ammonium chloride, ammonium hydroxide, ammonium acetate, ammonium dextran sulfate or triethyl ammonium dextran sulfate and combinations thereof.
  • the capture agent is ammonium sulfate or copper gluconate.
  • the concentration of the capture agent in the aqueous phase within the liposome is selected from 10 to 400mM, and can be 10mM, 15mM, 20mM, 25mM, 30mM, 35mM, 40mM, 45mM, 50mM, 55mM, 60mM, 65mM, 70mM, 75mM, 80mM, 85mM, 90mM, 95mM, 100mM, 105mM, 110mM, 115mM, 120mM, 125mM, 130mM, 135mM, 140mM, 145mM, 150mM, 155mM, 160mM, 165mM, 170mM, 175 mM, 180mM, 185mM, 190mM, 195mM, 200mM, 205mM, 210mM, 215mM, 220mM, 225mM, 230mM, 235mM, 240mM, 245mM, 250mM, 255
  • the pharmaceutical composition is a liposome comprising a compound of Formula I or a pharmaceutically acceptable salt thereof, a lipid and a capture agent, wherein the lipid comprises at least one phospholipid and a steroid.
  • the phospholipid is selected from dipalmitoylphosphatidylcholine (DPPC), distearoylphosphatidylcholine (DSPC), dioleoylphosphatidylcholine (DOPC), dierucylphospholipid Acylcholine (DEPC), dilauroylphosphatidylcholine (DLPC), hydrogenated soybean phosphatidylcholine (HSPC), distearoylphosphatidylethanolamine (DSPE) and dimyristoylphosphatidylcholine (DMPC) and their polyethylene glycol modified derivatives; the steroid is cholesterol.
  • DPPC dipalmitoylphosphatidylcholine
  • DSPC distearoylphosphatidylcholine
  • DOPC dioleoylphosphatidylcholine
  • DEPC dierucylphospholipid Acylcholine
  • DLPC dilauroylphosphatidylcholine
  • HSPC hydrogenated soybean phosphatidylcho
  • the liposome internal aqueous phase of the present disclosure also contains at least one pH regulator.
  • the pH regulator can be selected from amino acids such as arginine, histidine, glycine, ethylenediaminetetraacetic acid, Acids such as pentic acid, hydrochloric acid, phosphoric acid, malic acid or sulfuric acid; salts such as sodium salt, potassium salt and ammonium salt of the aforementioned acids; basic compounds (bases) such as ammonia, sodium hydroxide, potassium hydroxide, triethanolamine and the like. Examples include sodium hydroxide, hydrochloric acid, ammonia, phosphoric acid, malic acid, histidine, and triethanolamine.
  • the pH adjuster can be used in combination with two or more ammonium salts mentioned above.
  • the concentration of the pH regulator in the aqueous phase within the liposome is selected from 1 to 300mM, and can be 1mM, 2mM, 3mM, 4mM, 5mM, 6mM, 7mM, 8mM, 9mM, 10mM, 15mM, 1 35mM, 140mM, 145mM, 150mM, 155mM, 160mM, 165mM, 170mM, 175mM, 180mM, 185mM, 190mM, 195mM, 200mM, 205mM, 210mM, 215mM, 220mM, 225mM, 230mM, 235mM, 240mM, 245 mM, 250mM, 255mM, 260mM, 265mM, 270mM, 275mM, 280mM, 285mM, 290mM, 295mM, 300mM or any value between any two values, for example, it can be 1mM
  • the concentration of the compound of formula I or a pharmaceutically acceptable salt thereof is selected from 0.01 to 100 mg/mL, and can be 0.01 mg/mL, 0.05 mg/mL, 0.10 mg/mL, 0.15 mg/mL, 0.20 mg /mL, 0.25mg/mL, 0.30mg/mL, 0.35mg/mL, 0.40mg/mL, 0.45mg/mL, 0.50mg/mL, 0.55mg/mL, 0.60mg/mL, 0.65mg/mL, 0.70mg /mL, 0.75mg/mL, 0.80mg/mL, 0.85mg/mL, 0.90mg/mL, 0.95mg/mL, 1mg/mL, 2mg/mL, 3mg/mL, 4mg/mL, 5mg/mL, 6mg/ mL, 7mg/mL, 8mg/mL, 9mg/m
  • a solution containing electrolytes can be added to the liposome external phase to increase the ionic strength, thereby increasing the ionic strength.
  • the electrolyte (salt) contained in the liposome external phase is not particularly limited, and may be, for example, sodium chloride or potassium chloride, such as sodium chloride. Alternatively, physiological saline can also be used.
  • the liposome external phase such as a liposome dispersion may contain sugar, electrolytes, and/or amino acids or salts thereof, or may contain sugar, electrolytes, and amino acids or salts thereof.
  • the sugar is selected from sucrose or trehalose or glucose
  • the electrolyte is selected from physiological saline or sodium chloride or potassium chloride
  • the amino acid or its salt can be selected from histidine or its salt.
  • the liposome external phase contains sodium chloride and histidine or a salt thereof.
  • the concentration of the amino acid in the liposome external phase is selected from 1 to 300mM, and can be 1mM, 2mM, 3mM, 4mM, 5mM, 6mM, 7mM, 8mM, 9mM, 10mM, 15mM, 20mM, 25mM, 30mM, 35mM, 40mM, 45mM, 50mM, 55mM, 60mM, 65mM, 70mM, 75mM, 80mM, 85mM, 90mM, 95mM, 100mM, 105mM, 110mM, 115mM, 120mM, 125mM, 130mM, 135mM, 140mM ,145mM,150mM, 155mM, 160mM, 165mM, 170mM, 175mM, 180mM, 185mM, 190mM, 195mM, 200mM, 205mM, 210mM, 215mM, 220mM,
  • the pharmaceutical composition is a liposome comprising a compound of Formula I, or a pharmaceutically acceptable salt thereof, and a lipid, wherein the lipid comprises at least one phospholipid and a steroid, and the liposome
  • the inner aqueous phase contains a capture agent and a pH adjuster
  • the outer aqueous phase of the liposome contains one or more selected from sugars, electrolytes, and amino acids or salts thereof.
  • the pH of the internal aqueous phase of the liposomes of the present disclosure is 5-8; the pH of the external phase is 5-11.
  • the pH of the internal aqueous phase is 5-8; the pH of the external phase is 5-8.
  • the present disclosure also provides a method for preparing liposomes, which includes: preparing blank liposomes, and introducing the compound of formula I or a pharmaceutically acceptable salt thereof into the aqueous phase of the blank liposomes.
  • the present disclosure can use thin film dispersion method, reverse phase evaporation method, direct hydration method, surfactant removal method, ethanol injection method and modified ethanol injection method to prepare blank liposomes.
  • probe ultrasound, high-pressure homogenizer, microfluidizer, extruder and other devices can be used to obtain blank liposomes with the desired particle size.
  • various parameters (such as the amount of membrane components, temperature, etc.) during the preparation process of liposomes can be appropriately adjusted and determined according to the preparation method of liposomes or the composition, particle size, etc. of the target liposomes.
  • the replacement or dilution operation of the present disclosure can be performed by dialysis, centrifugation, gel filtration and other methods.
  • active substances can also be effectively encapsulated in the liposome internal aqueous phase.
  • Dialysis, centrifugation and gel filtration are all common operations in this field.
  • dialysis can be performed using a dialysis membrane.
  • the dialysis membrane can be made of polyethersulfone.
  • the pharmaceutical composition is a liquid preparation
  • the liposome composition obtained from the above introduction step can be directly used as the final liposome composition (pharmaceutical composition), or can be adjusted (substitution, etc.)
  • the liposome external phase of the obtained liposome composition is obtained, thereby obtaining the final liposome composition (pharmaceutical composition).
  • the pharmaceutical composition is a solid preparation
  • the liposome composition obtained from the above introduction step can be dried to obtain the final solid liposome composition (pharmaceutical composition).
  • Methods for drying the liposome composition may include, but are not limited to, freeze-drying and spray-drying.
  • the pharmaceutical composition is a solid preparation, and the pharmaceutical composition can be used as a liquid preparation by dissolving or suspending it in an appropriate solvent.
  • the solvent can be appropriately set according to the purpose of use of the liposome composition.
  • the solvent is preferably sterilized distilled water.
  • the present disclosure also provides a freeze-dried composition, which is obtained by freeze-drying the aforementioned pharmaceutical composition, or the aforementioned pharmaceutical composition is obtained after the freeze-dried composition is reconstituted with a liquid medium, wherein the liquid medium used for reconstitution is selected from the group consisting of physiological Saline, water for injection, glucose injection or glucose sodium chloride injection.
  • the present disclosure further provides the use of the aforementioned pharmaceutical composition, liposome, lyophilized composition or reconstituted solution in the preparation of a medicament for the treatment and/or prevention of a disease or disorder, the disease or disorder being cancer.
  • the disease or condition described in this disclosure is selected from the group consisting of brain cancer, thyroid cancer, head and neck cancer, nasopharyngeal cancer, throat cancer, oral cavity cancer, salivary gland cancer, esophageal cancer, gastric cancer, lung cancer, liver cancer, kidney cancer, pancreatic cancer, gallbladder cancer , cholangiocarcinoma, colorectal cancer, small bowel cancer, gastrointestinal stromal tumor, urothelial cancer, urethra cancer, bladder cancer, breast cancer, vaginal cancer, ovarian cancer, endometrial cancer, cervical cancer, fallopian tube cancer, testis Carcinoma, prostate cancer, hemangioma, leukemia, lymphoma, myeloma, skin cancer, lipoma, bone cancer, soft tissue sarcoma, neurofibroma, glioma, neuroblastoma, and glioblastoma.
  • the disease or disorder is selected from pancreatic cancer, colorectal cancer, and non-small cell lung cancer.
  • the pharmaceutical composition described in the present disclosure has good stability, high liposome encapsulation rate, large drug loading capacity, low liposome allergic reaction, and can be better used in clinical applications.
  • the "mass volume percentage” (w/v) mentioned in this disclosure refers to the mass (unit g) of the component contained in every 100 mL of liquid system, that is, g/100 mL.
  • the content of the compound of formula I or its pharmaceutically acceptable salt in the composition is 5 ⁇ 20 mg/mL
  • the content of the compound of formula I or its pharmaceutically acceptable salt in the composition is 4.75 to 21 mg/mL, which falls within the scope of the present disclosure
  • the compound of formula I or its pharmaceutically acceptable salt and the lipid weight ratio selection "From 1:1 to 1:100” means that the weight ratio of the compound of formula I or its pharmaceutically acceptable salt to the lipid is selected from 1:0.95 to 1:105, which all fall within the scope of the present disclosure.
  • mixing in and “mixing” in this disclosure are not limited to the order in which the components are added.
  • mixing A into B can mean that A is added to B, or it can be expressed that B is added to A.
  • Mixing A and B it can express the meaning of A added to B and mixed, and it can also express the meaning of B added to A mixed.
  • Liposomes refer to microscopic lipid membrane vesicles composed of a bilayer of phospholipids or any similar amphoteric lipids encapsulating an internal aqueous medium.
  • Liposomes of the present disclosure can be multilamellar vesicles (MLV), large unilamellar vesicles (LUV), small unilamellar vesicles. The size usually ranges from 30nm to 200nm.
  • MLV multilamellar vesicles
  • LUV large unilamellar vesicles
  • small unilamellar vesicles The size usually ranges from 30nm to 200nm.
  • Liposome membrane refers to the double layer of phospholipids that separates the internal aqueous medium from the external aqueous medium.
  • aqueous phase within the liposome refers to the aqueous region surrounded by the lipid bilayer of the liposome, and is used in the same meaning as the "internal water phase” and the "aqueous phase within the liposome".
  • liposome external phase refers to the area not surrounded by the lipid bilayer of the liposome (ie, the area other than the internal aqueous phase and lipid bilayer).
  • Blank liposomes refer to the aqueous phase within the liposomes that does not contain active ingredients to be loaded, such as the compound of Formula I of the present disclosure.
  • Encapsulation rate refers to the percentage of the encapsulated substance (such as a certain drug) in the liposome suspension to the total amount of the drug. It is an important indicator for quality control of liposomes and nanoparticles, reflecting the extent to which the drug is encapsulated by the carrier.
  • Encapsulation rate includes the measurement of percentage encapsulation rate and package volume.
  • General literature mainly examines the percentage encapsulation rate (Encapsulation percentage, EN%).
  • EN% (1-Cf/Ct) ⁇ 100%.
  • Cf is the amount of free drug
  • Ct is the total amount of drug in the liposome suspension.
  • the encapsulation efficiency can be measured using conventional methods for measuring the encapsulation efficiency, such as HPLC method.
  • the "polymer molecular weight” in this disclosure adopts the number average molecular weight, which refers to the statistical average of the molecular weights of all polymer chains in the sample, which is defined as follows, where M i represents the single chain molecular weight and N i represents the number of chains with corresponding molecular weights.
  • M n can be predicted from the aggregation mechanism and determined by measuring the number of molecules in a sample of a given mass; for example, colligative methods such as end-group analysis. If M n is used to characterize the molecular weight distribution, then there are an equal number of molecules distributed on both sides of M n .
  • particle size distribution refers to the particle size distribution in a liposome composition as measured by dynamic light scattering techniques well known to those skilled in the art, such as using Malvern Mastersizer TM 2000.
  • d(0.1) mentioned in this disclosure refers to the particle size corresponding to when the cumulative particle size distribution percentage of a sample reaches 10%.
  • d(0.5) refers to the particle size corresponding to when the cumulative particle size distribution percentage of a sample reaches 50%.
  • d(0.9) refers to the particle size corresponding to when the cumulative particle size distribution percentage of a sample reaches 90%.
  • Figure 1 is a cryo-scanning electron microscope (Cryo-TEM) picture of the liposome of Example 5;
  • Figure 2 is a cryo-scanning electron microscope (Cryo-TEM) picture of the liposome of Example 6.
  • the structure of the compound is determined by nuclear magnetic resonance (NMR) or/and mass spectrometry (MS). NMR shifts ( ⁇ ) are given in units of 10 -6 (ppm). NMR is measured using Bruker AVANCE-400 nuclear magnetic instrument or Bruker AVANCE NEO 500M.
  • the measurement solvents are deuterated dimethyl sulfoxide (DMSO-d 6 ), deuterated chloroform (CDCl 3 ), and deuterated methanol (CD 3 OD).
  • the internal standard is tetramethylsilane (TMS).
  • MS was measured using Agilent 1200/1290 DAD-6110/6120 Quadrupole MS liquid mass spectrometer (manufacturer: Agilent, MS model: 6110/6120 Quadrupole MS).
  • HPLC High-performance liquid chromatography
  • Chiral HPLC analysis was performed using an Agilent 1260 DAD high performance liquid chromatograph.
  • High-performance liquid phase preparation uses Waters 2545-2767, Waters 2767-SQ Detecor2, Shimadzu LC-20AP and Gilson GX-281 preparative chromatographs.
  • Chiral preparation uses Shimadzu LC-20AP preparative chromatograph.
  • CombiFlash rapid preparation instrument uses Combiflash Rf200 (TELEDYNE ISCO).
  • Thin layer chromatography silica gel plates use Yantai Huanghai HSGF254 or Qingdao GF254 silica gel plates.
  • the specifications of silica gel plates used in thin layer chromatography (TLC) are 0.15mm ⁇ 0.2mm.
  • the specifications used for thin layer chromatography separation and purification products are 0.4mm. ⁇ 0.5mm.
  • Silica gel column chromatography generally uses Yantai Huanghai Silica Gel 200 ⁇ 300 mesh silica gel as the carrier.
  • the average kinase inhibition rate and IC 50 value were measured using NovoStar microplate reader (BMG Company, Germany).
  • the known starting materials of the present invention can be synthesized by methods known in the art, or can be purchased from ABCR GmbH&Co.KG, Acros Organics, Aldrich Chemical Company, Shaoyuan Chemical Technology (Accela ChemBio Inc), Darui Chemical companies.
  • Argon atmosphere or nitrogen atmosphere means that the reaction bottle is connected to an argon or nitrogen balloon with a volume of about 1L.
  • the hydrogen atmosphere refers to the reaction bottle connected to a hydrogen balloon with a volume of about 1L.
  • the pressurized hydrogenation reaction uses Parr 3916EKX hydrogenator and Clear Blue QL-500 hydrogen generator or HC2-SS hydrogenator.
  • the hydrogenation reaction is usually evacuated, filled with hydrogen, and repeated three times.
  • the microwave reaction uses CEM Discover-S 908860 microwave reactor.
  • the solution refers to an aqueous solution.
  • the reaction temperature is room temperature, which is 20°C to 30°C.
  • the reaction progress in the embodiment is monitored by thin layer chromatography (TLC).
  • TLC thin layer chromatography
  • the developing agent used in the reaction, the column chromatography eluent system and the thin layer chromatography developing agent system used to purify the compound include: A: Dichloromethane/methanol system, B: n-hexane/ethyl acetate, the volume ratio of the solvent is adjusted according to the polarity of the compound, and a small amount of alkaline or acidic reagents such as triethylamine and acetic acid can also be added for adjustment.
  • the relative stereochemistry of these compounds was identified by NMR studies and/or X-ray diffraction. In these cases, the compounds are identified using the prefix "rel” followed by R/S nomenclature, where the R/S only provides relative stereochemistry information and does not indicate absolute stereochemistry. For example, express The 1:1 mixture is the racemate.
  • HPLC analysis retention time 1.02 minutes, purity: 98.5% (Column: ACQUITY C18, 1.7 ⁇ m, 2.1*50mm; mobile phase: water (10mM ammonium bicarbonate), acetonitrile, gradient ratio: acetonitrile 10%-95%).
  • Dissolve compound 2a (1.8g, 9.94mmol) in tetrahydrofuran (50mL), add 20mL of 2M sodium bistrimethylsilylamide tetrahydrofuran solution under ice bath, stir and react for 0.5 hours, add di-tert-butyl dicarbonate (6.5 g, 29.7 mmol), stir for 14 hours, add saturated ammonium chloride aqueous solution to the reaction solution to quench, extract with ethyl acetate (50 mL The mixture was concentrated under reduced pressure, and the residue was purified with eluent system B to obtain the title compound 3b (1 g, yield: 26.3%). The product was directly used in the next reaction without purification.
  • Dissolve compound 3b (1g, 2.62mmol) in ethyl acetate (8mL), add 3mL of 4M dioxane hydrochloride solution, stir the reaction for 2 hours, and adjust the pH to neutral with 4M sodium hydroxide aqueous solution in an ice bath. , extracted with ethyl acetate (10 mL rate: 67.8%).
  • the crude compound 3d (835 mg, 1.77 mmol) was dissolved in 7M ammonia-methanol solution (10 mL), and the reaction was stirred for 1 hour. The reaction solution was concentrated under reduced pressure. Methyl tert-butyl ether (10 mL) was added to the residue, and stirred for 0.5 hours. After filtering and drying the filter cake, the crude title compound 3e (400 mg, yield: 89.9%) was obtained. The product was directly used in the next reaction without purification.
  • reaction solution was concentrated under reduced pressure, and the residue was analyzed by high-performance liquid chromatography (Waters-2545, column: SharpSil-T C18, 30*150mm, 5 ⁇ m; mobile phase: water phase (10mmol/L ammonium bicarbonate) and acetonitrile, gradient Proportion: acetonitrile 38%-45%, flow rate: 30 mL/min) and purified to obtain a 1:1 mixture of diastereomers of the title compound 3p-1 and 3p-2 (620 mg, yield: 32.6%).
  • high-performance liquid chromatography Waters-2545, column: SharpSil-T C18, 30*150mm, 5 ⁇ m; mobile phase: water phase (10mmol/L ammonium bicarbonate) and acetonitrile, gradient Proportion: acetonitrile 38%-45%, flow rate: 30 mL/min
  • the diastereomeric mixture of compounds 3p-1 and 3p-2 was resolved by chiral column (Shimadzu LC-20AP, column: DAICEL 25*250mm, 10 ⁇ m; mobile phase A: n-hexane, mobile phase B: ethanol (0.1% 7M NH3in MeOH)) and, gradient ratio: A:B: 30:70, flow rate: 30mL/min) to obtain the title compound 3p -1 (50 mg, yield: 35.7%) and 3p-2 (50 mg, yield: 35.7%).
  • Chiral HPLC analysis retention time 9.85 minutes, purity: 99% (Column: DAICEL 250*4.6mm, 5 ⁇ m; mobile phase: n-hexane and ethanol (containing 0.2% diethylamine), flow rate: 1.0mL/min).
  • Chiral HPLC analysis retention time 16.0 minutes, purity: 99% (Column: DAICEL 250*4.6mm, 5 ⁇ m; mobile phase: n-hexane and ethanol (containing 0.2% diethylamine), flow rate: 1.0mL/min).
  • the diastereomeric mixture of compounds 4p-1 and 4p-2 was resolved by chiral column (Shimadzu LC-20AP, column: DAICEL 25*250mm, 10 ⁇ m; mobile phase A: n-hexane, mobile phase B: ethanol (0.1% 7M NH3in MeOH)) and, gradient ratio: A:B: 40:60, flow rate: 30mL/min) to obtain the title compound 4p -1 (26 mg, yield: 43.3%) and 4p-2 (26 mg, yield: 43.3%).
  • Chiral HPLC analysis retention time 13.8 minutes, purity: 99% (Column: DAICEL 250*4.6mm, 5 ⁇ m; mobile phase: n-hexane and ethanol (containing 0.2% diethylamine), flow rate: 1.0mL/min).
  • Test Example 1 Experimental biological evaluation of ERK phosphorylation inhibition in AGS cells (HTRF method)
  • This experiment detects the inhibitory effect of the compound represented by formula I on cellular ERK phosphorylation, and evaluates the inhibitory effect of the compound on the KRAS target according to the IC 50 size.
  • AGS cells (Nanjing Kebai, CBP60476) were cultured in RPMI1640 (Hyclone, SH30809.01) complete medium containing 10% fetal calf serum. On the first day of the experiment, AGS cells were seeded in a 96-well plate at a density of 40,000 cells/well using complete culture medium, with 190 ⁇ L of cell suspension in each well, and cultured overnight in a 37°C, 5% CO 2 cell incubator.
  • lysis buffer lysis buffer, Cisbio, 64KL1FDF
  • blocking reagent Cisbio, 64KB1AAC
  • 4 ⁇ L of premixed phosphorylated ERK1/2 antibody solution to the two plates.
  • Cisbio, 64AERPEG premixed phosphorylated ERK1/2 antibody solution
  • the microplate was sealed with sealing film, centrifuged in a microplate centrifuge for 1 minute, and incubated overnight at room temperature in the dark.
  • the IC 50 value of the inhibitory activity of the compound was calculated based on the compound concentration and the ratio of phosphorylated ERK/total ERK using Graphpad Prism software. The results are shown in Table 1 below.
  • Test Example 2 Experimental biological evaluation of 3D proliferation inhibition of GP2d and AGS cells
  • GP2d cells (Nanjing Kebai, CBP60010) were cultured in complete medium, that is, DMEM/high glucose medium (Hyclone, SH30243.01) containing 10% fetal calf serum (Corning, 35-076-CV).
  • GP2d cells were seeded in a 96-well low-adsorption plate (Corning, CLS7007-24EA) using complete culture medium at a density of 1,000 cells/well, with 90 ⁇ L of cell suspension in each well, centrifuged at room temperature at 2,000 rpm for 5 minutes, and then placed for 37 °C, 5% CO2 cell culture incubator overnight.
  • AGS cells (Nanjing Kebai, CBP60476) were cultured in complete medium, that is, RPMI1640 medium (Hyclone, SH30809.01) containing 10% fetal calf serum (Corning, 35-076-CV).
  • complete medium that is, RPMI1640 medium (Hyclone, SH30809.01) containing 10% fetal calf serum (Corning, 35-076-CV).
  • RPMI1640 medium Hyclone, SH30809.01
  • fetal calf serum (Corning, 35-076-CV).
  • AGS cells were seeded in a 96-well low-adsorption plate (Corning, CLS7007-24EA) using complete culture medium at a density of 1,000 cells/well, with 90 ⁇ L of cell suspension in each well, centrifuged at 2,000 rpm for 5 minutes at room temperature, and then placed for 37 °C, 5% CO2 cell culture incubator overnight.
  • the next day add 10 ⁇ L of a gradient dilution of the compound to be tested prepared in complete culture medium to each well.
  • the final concentration of the compound for GP2d cells is 9 concentration points of 3-fold gradient dilution starting from 1 ⁇ M.
  • the final concentration of the compound for AGS cells is from 1 ⁇ M.
  • a blank control containing 0.5% DMSO was set.
  • the well plate was placed in a 37°C, 5% CO2 cell incubator for 120 hours.
  • 3D reagent (Promega, G9682), shake at room temperature for 25 minutes, mix by pipetting, and transfer 50 ⁇ L to a white 96-well plate (PE, 6005290) with opaque bottom, using a multifunctional microplate reader (PerkinElmer, ENVISION ) to read the luminescence signal value.
  • the compound represented by formula I has a good inhibitory effect on the 3D proliferation of AGS and GP2d cells.
  • Test Example 3 SPR method to detect the affinity of compounds to KRAS protein isoform G12D or WT
  • Biotinylated Avi-KRAS-WT or Avi-KRAS-G12D was first diluted to 20 ⁇ g/mL with 1 ⁇ HBS-P+ (Cat.#BR1006-71) buffer containing 100mM MgCl, and then flowed through SA (Cat .#BR1005-31) Biosensor chip channel 2, lasting 420s, obtaining a coupling level of approximately 5000-7000RU. Then compound samples were injected sequentially from high to low for 120 s, and then dissociated for 720 s. The experiment adopts single cycle kinetic mode.
  • the Biacore 8K instrument detects the reaction signal in real time to obtain the binding and dissociation curve. After the test, Biacore 8K evaluation software was used for data analysis, and a 1:1 model was used to fit the data and obtain affinity data. The results are shown in Table 3 below.
  • the compound represented by formula I has good affinity to KRAS protein isoform G12D or WT.

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Abstract

本公开涉及一种包含KRAS G12D抑制剂的药物组合物。具体而言，本公开涉及一种药物组合物，其包含式I化合物或其可药用盐和脂质。该药物组合物可以更好地应用于临床。

Description

一种包含KRAS G12D抑制剂的药物组合物 技术领域
本公开属于药物制剂领域，具体涉及一种包含KRAS G12D抑制剂的药物组合物及其制备方法。
背景技术
RAS是在肿瘤中突变率最高的致癌基因之一，约30％的人类恶性肿瘤与RAS基因的突变有关。RAS家族包括KRAS，NRAS和HRAS，其中KRAS突变最为常见，约占85％。KRAS突变常见于实体瘤，在人类三大致命性癌症—肺癌(17％)、结直肠癌(33％)和胰腺癌(61％)中均存在高频突变。在KRAS的基因突变中，97％是第12号或者第13号氨基酸残基发生了突变，其中G12D是一个重要突变。对欧美人群的数据分析显示：在胰腺癌、结直肠癌及非小细胞肺癌中，G12D突变分别占病人的36％、12％和4％。
KRAS被激活以后，通过以RAF-MEK-ERK，PI3K-AKT-mTOR及TIAM1-RAc为代表的众多下游信号通路，调控细胞增殖、存活、迁移及代谢等多个方面的功能。KRAS基因突变后，蛋白持续处于活化状态，导致下游信号通路持续激活而促进肿瘤发生。
由于KRAS蛋白表面缺乏传统意义上的小分子结合位点，并与鸟苷酸有着超高亲和力而极难被抑制，长久以来被认为是不可成药的药物靶点。但基于KRAS异常激活在癌症进展中的重要性和普遍性，KRAS一直并仍然是药物开发非常关注的靶点。目前除了KRAS G12C抑制剂以外，仍缺乏对其他突变有效的KRAS抑制剂，使得大部分KRAS突变的病人依然无药可治。G12D，作为一个在多种肿瘤中广泛高表达的突变体，开发针对它的抑制剂有着重要的临床意义。
WO2022268051提供了一种新的KRAS G12D抑制剂化合物(式I)，该化合物具有较好的药学活性。
发明内容
本公开的目的在于提供一种包含式I化合物或其可药用盐的药物组合物。
本公开一方面提供了一种药物组合物，其包含式I化合物或其可药用盐和脂质，
在一些实施方案中，所述脂质包含至少一种磷脂。
在一些实施方案中，本公开所述的药物组合物为脂质体。
本公开另一方面提供了一种包含式I化合物或其可药用盐的脂质体。
本公开所述的脂质可包括完全中性或带负电荷的磷脂。术语“磷脂”指含有至少一个磷基的疏水分子，其可是天然或合成的。例如，磷脂可包含含磷基团及任选被OH、COOH、氧代、胺或者取代或未取代的芳基取代的饱和或不饱和的烷基。磷脂类在其无环(acylic)链的长度和不饱和度方面互相不同。
在一些实施方案中，所述磷脂包括磷脂酰胆碱、磷脂酰乙醇胺、磷脂酰甘油、磷脂酰丝氨酸、磷脂酸、磷脂酰肌醇中的一种或多种。术语“磷脂酰胆碱”指磷脂酰胆碱及其衍生物。适合用于本公开的磷脂的示例包括二棕榈酰磷脂酰胆碱(DPPC)、二硬脂酰磷脂酰胆碱(DSPC)、二肉豆蔻酰磷脂酰胆碱(DMPC)、1-棕榈酰-2-亚油酰-sn-甘油-3-磷脂酰胆碱(PLPC)、二油酰磷脂酰胆碱(DOPC)、二芥酰磷脂酰胆碱(DEPC)、蛋黄磷脂酰胆碱(EPC)、二月桂酰磷脂酰胆碱(DLPC)、氢化大豆磷脂酰胆碱(HSPC)、l-肉豆蔻酰-2-棕榈酰磷脂酰胆碱(MPPC)、l-棕榈酰-2-肉豆蔻酰磷脂酰胆碱(PMPC)、l-棕榈酰-2-硬脂酰磷脂酰胆碱(PSPC)、l-硬脂酰-2-棕榈酰磷脂酰胆碱(SPPC)、棕榈酰油酰磷脂酰胆碱(POPC)、溶血磷脂酰胆碱、二亚油酰磷脂酰胆碱、二硬脂酰磷脂酰乙醇胺(DSPE)、二肉豆蔻酰磷脂酰乙醇胺(DMPE)、二棕榈酰磷脂酰乙醇胺(DPPE)、二油酰基磷脂酰甘油(DOPG)、二肉豆蔻酰磷脂酰甘油(DMPG)、二硬脂酰基磷脂酰甘油(DSPG)、二棕榈酰基甘油磷酸甘油(DPPG)、二棕榈酰基磷脂酰丝氨酸(DPPS)、1，2-二油酰基-sn-甘油-3-磷脂酰丝氨酸(DOPS)、二肉豆蔻酰基磷脂酰丝氨酸(DMPS)、二硬脂酰基磷脂酰丝氨酸(DSPS)、二棕榈酰基磷脂酸(DPPA)、1，2-二油酰基-sn-甘油-3-磷脂酸(DOPA)、二肉豆蔻酰基磷脂酸(DMPA)、二硬脂酰基磷脂酸(DSPA)、二棕榈酰基磷脂酰肌醇(DPPI)、1，2-二油酰基-sn-甘油-3-磷脂酰肌醇(DOPI)、二肉豆蔻酰基 磷脂酰肌醇(DMPI)、二硬脂酰基磷脂酰肌醇(DSPI)，以及它们的衍生物(例如亲水性聚合物修饰的衍生物)。
所述亲水性聚合物修饰的磷脂包括但不限于聚乙二醇修饰的二硬脂酰磷脂酰乙醇胺(PEG-DSPE)、聚乙二醇修饰的二硬脂酰磷脂酰甘油(PEG-DSPG)、聚乙二醇修饰的胆固醇(PEG-CHOL)。PEG的分子量为200-10000道尔顿，例如可以是1000-5000道尔顿，或者2000-5000道尔顿。
在一些实施方案中，本公开中所述的磷脂选自二棕榈酰磷脂酰胆碱(DPPC)、二硬脂酰磷脂酰胆碱(DSPC)、二油酰磷脂酰胆碱(DOPC)、二芥酰磷脂酰胆碱(DEPC)、二月桂酰磷脂酰胆碱(DLPC)、氢化大豆磷脂酰胆碱(HSPC)、二硬脂酰磷脂酰乙醇胺(DSPE)和二肉豆蔻酰磷脂酰胆碱(DMPC)以及它们的聚乙二醇修饰的衍生物。
在一些实施方案中，所述脂质体含有聚乙二醇修饰的二硬脂酰磷脂酰乙醇胺。在一些实施方案中，所述聚乙二醇修饰的二硬脂酰磷脂酰乙醇胺中聚乙二醇分子量为200-10000道尔顿。在一些实施方案中，所述聚乙二醇修饰的二硬脂酰磷脂酰乙醇胺中聚乙二醇分子量为1000-5000道尔顿。在一些实施方案中，所述聚乙二醇修饰的二硬脂酰磷脂酰乙醇胺中聚乙二醇分子量为2000-5000道尔顿。
在一些实施方案中，所述聚乙二醇修饰的二硬脂酰磷脂酰乙醇胺选自聚乙二醇2000-二硬脂酰磷脂酰乙醇胺(mPEG2000-DSPE)。
在一些实施方案中，相对于总脂质混合物的量，所述磷脂摩尔含量选自10～80％，包括但不限于10％、11％、12％、13％、14％、15％、16％、17％、18％、19％、20％、21％、22％、23％、24％、25％、26％、27％、28％、29％、30％、31％、32％、33％、34％、35％、36％、37％、38％、39％、40％、41％、42％、43％、44％、45％、46％、47％、48％、49％、50％、51％、52％、53％、54％、55％、56％、57％、58％、59％、60％、61％、62％、63％、64％、65％、66％、67％、68％、69％、70％、71％、72％、73％、74％、75％、76％、77％、78％、79％、80％或任意两数值间任意值，例如可以是30～70％。
本公开还可包含其他中性脂质、阳离子脂质和/或阴离子脂质。
可用于本公开的其他中性脂质的示例包括：类固醇如胆固醇及其衍生物、卵磷脂、大豆磷脂、脑磷脂、鞘磷脂、氢化大豆磷脂中的一种或多种。在一些实施方案中，相对于总脂质混合物的量，类固醇的摩尔百分比不超过90％。
在一些实施方案中，所述的脂质包含至少一种磷脂以及胆固醇。相对于总脂质混合物的量，胆固醇的摩尔百分比可以是0.1％-90％，例如可以是10％-80％，或者20％-70％。
在一些实施方案中，所述的磷脂选自二棕榈酰磷脂酰胆碱(DPPC)、二硬脂酰磷脂酰胆碱(DSPC)、二油酰磷脂酰胆碱(DOPC)、二芥酰磷脂酰胆碱(DEPC)、二 月桂酰磷脂酰胆碱(DLPC)、氢化大豆磷脂酰胆碱(HSPC)、二硬脂酰磷脂酰乙醇胺(DSPE)和二肉豆蔻酰磷脂酰胆碱(DMPC)以及它们的聚乙二醇修饰的衍生物。
在一些实施方案中，所述脂质包含氢化大豆磷脂酰胆碱、胆固醇及聚乙二醇2000-二硬脂酰磷脂酰乙醇胺。
在一些实施方案中，所述亲水性聚合物修饰的磷脂衍生物含量(相对全部脂质体膜成分，摩尔分数)选自0.1～50％，包括但不限于0.1％、0.2％、0.3％、0.4％、0.5％、0.6％、0.7％、0.8％、0.9％、1％、2％、3％、4％、5％、6％、7％、8％、9％、10％、11％、12％、13％、14％、15％、16％、17％、18％、19％、20％、21％、22％、23％、24％、25％、26％、27％、28％、29％、30％、31％、32％、33％、34％、35％、36％、37％、38％、39％、40％、41％、42％、43％、44％、45％、46％、47％、48％、49％、50％或任意两数值间任意值，例如可以是0.5～20％。
在一些实施方案中，所述的药物组合物还包含捕获剂。在一些实施方案中，所述的脂质体内水相中包含捕获剂。
所述脂质体的制备方法可采用主动载药法。主动载药法为通过多原子离子梯度(polyatomic ion-gradient)将治疗剂从外部介质跨越脂质体的双层膜传输至内部水性空间的药物装载法。这些梯度通过在脂质体的内部水性空间中包埋至少一多原子离子作为捕获剂，并以例如，纯水、蔗糖溶液(sucrose solution)或生理盐水等具有较低多原子离子浓度的额外介质通过如管柱分离(column separation)、透析(dialysis)或离心(centrifugation)等已知技术置换脂质体的外部介质而产生。在脂质体的内部水性空间及外部介质之间制造多原子离子梯度以将治疗剂捕获于脂质体的内部水性空间。
本公开所述的捕获剂包括但不限于硫酸盐、亚硫酸盐、葡萄糖酸盐、磷酸盐、磷酸氢盐、钼酸盐、碳酸盐及硝酸盐等。例示性的捕获剂包含但不限于硫酸铵、磷酸铵、钼酸铵、蔗糖八硫酸酯铵、蔗糖八硫酸酯三乙铵、葡萄糖酸铜、乙基硫酸铵、乙二胺四乙酸铵盐、氯化铵、氢氧化铵、乙酸铵、葡聚糖硫酸铵或三乙基葡聚糖硫酸铵及其组合。
在一些实施方案中，所述捕获剂为硫酸铵或葡萄糖酸铜。
在一些实施方案中，所述脂质体内水相中捕获剂的浓度选自10～400mM，可以为10mM、15mM、20mM、25mM、30mM、35mM、40mM、45mM、50mM、55mM、60mM、65mM、70mM、75mM、80mM、85mM、90mM、95mM、100mM、105mM、110mM、115mM、120mM、125mM、130mM、135mM、140mM、145mM、150mM、155mM、160mM、165mM、170mM、175mM、180mM、185mM、190mM、195mM、200mM、205mM、210mM、215mM、220mM、225mM、230mM、235mM、240mM、245mM、250mM、255mM、260mM、265mM、270mM、275mM、280mM、285mM、290mM、295mM、 300mM、305mM、310mM、315mM、320mM、325mM、330mM、335mM、340mM、345mM、350mM、355mM、360mM、365mM、370mM、375mM、380mM、385mM、390mM、395mM、400mM或任意两数值间任意值，例如可以是20～400mM。
在一些实施方案中，所述药物组合物为脂质体，其包含式I化合物或其可药用盐，脂质和捕获剂，其中所述脂质包含至少一种磷脂和类固醇。
在一些实施方案中，所述的磷脂选自二棕榈酰磷脂酰胆碱(DPPC)、二硬脂酰磷脂酰胆碱(DSPC)、二油酰磷脂酰胆碱(DOPC)、二芥酰磷脂酰胆碱(DEPC)、二月桂酰磷脂酰胆碱(DLPC)、氢化大豆磷脂酰胆碱(HSPC)、二硬脂酰磷脂酰乙醇胺(DSPE)和二肉豆蔻酰磷脂酰胆碱(DMPC)以及它们的聚乙二醇修饰的衍生物；所述类固醇为胆固醇。
在一些实施方案中，本公开所述的脂质体内水相中还含有至少一种pH调节剂，pH调节剂可以选自精氨酸、组氨酸、甘氨酸等氨基酸，乙二胺四乙酸、喷替酸、盐酸、磷酸、苹果酸或硫酸等酸；前述酸的钠盐、钾盐、铵盐等盐；氨水、氢氧化钠、氢氧化钾、三乙醇胺等碱性化合物(碱)等。例如可以是氢氧化钠、盐酸、氨水、磷酸、苹果酸、组氨酸、三乙醇胺。另外，pH调节剂可以组合前述两种以上的铵盐使用。
在一些实施方案中，所述脂质体内水相中的pH调节剂的浓度选自1～300mM，可以为1mM、2mM、3mM、4mM、5mM、6mM、7mM、8mM、9mM、10mM、15mM、20mM、25mM、30mM、35mM、40mM、45mM、50mM、55mM、60mM、65mM、70mM、75mM、80mM、85mM、90mM、95mM、100mM、105mM、110mM、115mM、120mM、125mM、130mM、135mM、140mM、145mM、150mM、155mM、160mM、165mM、170mM、175mM、180mM、185mM、190mM、195mM、200mM、205mM、210mM、215mM、220mM、225mM、230mM、235mM、240mM、245mM、250mM、255mM、260mM、265mM、270mM、275mM、280mM、285mM、290mM、295mM、300mM或任意两数值间任意值，例如可以是20～300mM。
在一些实施方案中，所述式I化合物或其可药用盐的浓度选自0.01～100mg/mL，可以0.01mg/mL、0.05mg/mL、0.10mg/mL、0.15mg/mL、0.20mg/mL、0.25mg/mL、0.30mg/mL、0.35mg/mL、0.40mg/mL、0.45mg/mL、0.50mg/mL、0.55mg/mL、0.60mg/mL、0.65mg/mL、0.70mg/mL、0.75mg/mL、0.80mg/mL、0.85mg/mL、0.90mg/mL、0.95mg/mL、1mg/mL、2mg/mL、3mg/mL、4mg/mL、5mg/mL、6mg/mL、7mg/mL、8mg/mL、9mg/mL、10mg/mL、15mg/mL、20mg/mL、25mg/mL、30mg/mL、35mg/mL、40mg/mL、45mg/mL、50mg/mL、55mg/mL、60mg/mL、65mg/mL、70mg/mL、75mg/mL、 80mg/mL、85mg/mL、90mg/mL、95mg/mL、100mg/mL或任意两数值间任意值，例如可以是0.05～10mg/mL。
另一方面，为了改善式I化合物或其可药用盐在脂质体中的包封率，可以向脂质体外相中添加含有电解质的溶液(盐类溶液)使离子强度增大，由此增加包封率。脂质体外相中含有的电解质(盐类)没有特别限定，例如可以是为氯化钠、氯化钾，例如为氯化钠。另外，也可以使用生理盐水。另外，作为脂质体分散液等的脂质体外相，可以含有糖、电解质、及/或氨基酸或其盐，也可以含有糖或电解质、及氨基酸或其盐。作为示例，糖选自蔗糖或海藻糖或葡萄糖，电解质选自生理盐水或氯化钠或氯化钾，氨基酸或其盐可以选自组氨酸或其盐。
在一些实施方案中，所述脂质体外相中含有氯化钠和组氨酸或其盐。
在一些实施方案中，脂质体外相中所述氨基酸的浓度选自1～300mM，可以为1mM、2mM、3mM、4mM、5mM、6mM、7mM、8mM、9mM、10mM、15mM、20mM、25mM、30mM、35mM、40mM、45mM、50mM、55mM、60mM、65mM、70mM、75mM、80mM、85mM、90mM、95mM、100mM、105mM、110mM、115mM、120mM、125mM、130mM、135mM、140mM、145mM、150mM、155mM、160mM、165mM、170mM、175mM、180mM、185mM、190mM、195mM、200mM、205mM、210mM、215mM、220mM、225mM、230mM、235mM、240mM、245mM、250mM、255mM、260mM、265mM、270mM、275mM、280mM、285mM、290mM、295mM、300mM或任意两数值间任意值，例如可以是5-200mM。
在一些实施方案中，所述药物组合物为脂质体，其包含式I化合物或其可药用盐和脂质，其中所述脂质包含至少一种磷脂和类固醇，所述脂质体的内水相包含捕获剂和pH调节剂，所述脂质体的外水相包含选自糖、电解质和氨基酸或其盐中的一种或多种。
在一些实施方案中，本公开所述的脂质体的内水相pH为5-8；外相pH为5-11。优选内水相pH为5-8；外相pH为5-8。
本公开还提供一种制备脂质体的方法，包括：制备空白脂质体，以及将式I化合物或其可药用盐导入空白脂质体内水相的步骤。
本公开可采用薄膜分散法、逆相蒸发法、直接水化法、表面活性剂去除法、乙醇注入法以及改良乙醇注入法制备空白脂质体。
在一些实施方案中，可采用探头超声、高压均质机、微射流仪、挤出仪等装置处理获得所需粒径大小空白脂质体。
另外，脂质体的制备过程中各种参数(如膜成分的量、温度等)可以根据脂质体的制备方法或目标的脂质体的组成、粒径等适当调整确定。
另一方面，本公开置换或稀释操作可以通过透析、离心分离、凝胶过滤等方法进行。另外，通过置换或稀释脂质体外相，也可以有效地将活性物质包封在脂质体内水相中。
透析、离心分离和凝胶过滤均为本领域常规操作，例如透析可以使用透析膜进行。透析膜可选用聚醚砜材质的膜。
在一些实施方案中，药物组合物为液体制剂，可以将由上述导入的步骤得到的脂质体组合物可以直接作为最终的脂质体组合物(药物组合物)，或可以通过调节(置换等)得到的脂质体组合物的脂质体外相，从而获得最终的脂质体组合物(药物组合物)。
在另一些实施方案中，药物组合物为固体制剂，可以通过对由上述导入步骤得到的脂质体组合物进行干燥，作为最终的固态脂质体组合物(药物组合物)。对脂质体组合物进行干燥的方法，可以包括但不限于冻干及喷雾干燥等。
另一方面，药物组合物为固体制剂，药物组合物可以通过溶解或混悬在适当的溶剂中，作为液体制剂使用。溶剂可以根据脂质体组合物的使用目的等适当设定，例如药物组合物作为注射剂时，溶剂优选为灭菌蒸馏水。
本公开还提供一种冻干组合物，其由前述药物组合物冷冻干燥得到，或者所述冻干组合物经液体介质复溶后得到前述药物组合物，其中复溶使用的液体介质选自生理盐水、注射用水、葡萄糖注射液或葡萄糖氯化钠注射液。
本公开进一步提供前述的药物组合物、脂质体、冻干组合物或复溶溶液在制备用于治疗和/或预防疾病或病症的药物中的用途，所述的疾病或病症为癌症。
本公开中所述的疾病或病症选自脑癌、甲状腺癌、头颈癌、鼻咽癌、咽喉癌、口腔癌、唾液腺癌、食道癌、胃癌、肺癌、肝癌、肾癌、胰腺癌、胆囊癌、胆管癌、结直肠癌、小肠癌、胃肠道间质瘤、尿路上皮癌、尿道癌、膀胱癌、乳腺癌、阴道癌、卵巢癌、子宫内膜癌、宫颈癌、输卵管癌、睾丸癌、前列腺癌、血管瘤、白血病、淋巴瘤、骨髓瘤、皮肤癌、脂肪瘤、骨癌、软组织肉瘤、神经纤维瘤、神经胶质瘤、成神经细胞瘤和胶质母细胞瘤。
在一些实施方案中，所述的疾病或病症选自胰腺癌、结直肠癌和非小细胞肺癌。
本公开所述的药物组合物具有良好的稳定性，脂质体包封率高，载药量大，脂质体过敏反应低，可更好地应用于临床。
本公开所述的“质量体积百分比”(w/v)是指每100mL液体体系中含有所述成分的质量(单位g)，即g/100mL。
本公开所述的各种物质的含量(包括百分含量)及相互之间的比例，均允许存在±5％的误差，例如“组合物中式I化合物或其可药用盐的含量是5～20mg/mL”意味着组合物中式I化合物或其可药用盐含量为4.75～21mg/mL均属于本公开的范围；“所述的式I化合物或其可药用盐与所述的脂质的重量比选 自1:1～1:100”意味着所述的式I化合物或其可药用盐与所述的脂质的重量比选自1:0.95～1:105均属于本公开的范围。
本公开的“混入”、“混合”表示的意思是不限定组分的加入顺序，例如将A混入B，可以表达A加入B的意思，也可以表达B加入A的意思，将A和B混合，可以表达A加入B混合的意思，也可以表达B加入A混合的意思。
脂质体，是指有双层磷脂或任何类似的封装内部水性介质的两性脂质组成的显微脂质膜泡。本公开的脂质体可以是多层膜泡(MLV)、大单层膜泡(LUV)、小单层膜泡。尺寸通常从30nm～200nm。另外，对本公开中脂质体膜结构没有具体限制。
脂质体膜，是指分离内部水性介质与外部水性介质的双层磷脂。
脂质体内水相，是指被脂质体的脂质双层包围的水性区域，以“内水相”及“脂质体内水相”的相同含义进行使用。在脂质体分散在液体中时，“脂质体外相”是指没有被脂质体的脂质双层包围的区域(即除内水相及脂质双层之外的区域)。
空白脂质体，是指脂质体内水相中不含有待装载的活性成分如本公开式I化合物。
包封率，是指被包裹物质(如某药物)在脂质体悬液中占药物总量的百分量。它是脂质体和纳米粒质量控制的一个重要的指标，反映了药物被载体包封的程度。
包封率包括百分包封率和包裹容积的测定，一般文献主要考察百分包封率(Encapsulation percentage,EN％)。其表达式是EN％＝(1－Cf/Ct)×100％。式中Cf为游离药物的量；Ct为脂质体悬液中药物的总量。
可采用常规的包封率的测定方法测定包封率，例如HPLC法。
本公开中“聚合物分子量”采用数均分子量，是指样品中所有聚合链分子量的统计平均值，其定义如下其中Mi代表单链分子量，Ni代表具有相应分子量的链数目。Mn可以通过聚合机制来进行预测，并通过测定给定质量样品中的分子数量来确定；例如端基分析等依数性方法。如果用Mn来表征分子量分布，则Mn两侧分布有同等数量的分子。
术语“粒径分布”或“PSD”指的是通过本领域的技术人员熟知的动态光散射技术例如使用Malvern MastersizerTM2000所测量的脂质体组合物中的粒径分布。本公开所述的“d(0.1)”是指一个样品的累计粒度分布百分数达到10％时所对应的粒径。“d(0.5)”是指一个样品的累计粒度分布百分数达到50％时所对应的粒径。“d(0.9)”是指一个样品的累计粒度分布百分数达到90％时所对应的粒径。
附图说明
图1为实施例5脂质体冷冻扫描电镜(Cryo-TEM)图；
图2为实施例6脂质体冷冻扫描电镜(Cryo-TEM)图。
具体实施方式
通过以下实施例进一步详细说明本公开。这些实施例仅用于说明性目的，而并不用于限制本公开的范围。本公开全文引用PCT/CN2022/100016中的内容。
化合物的结构是通过核磁共振(NMR)或/和质谱(MS)来确定的。NMR位移(δ)以10-6(ppm)的单位给出。NMR的测定是用Bruker AVANCE-400核磁仪或Bruker AVANCE NEO 500M，测定溶剂为氘代二甲基亚砜(DMSO-d6)、氘代氯仿(CDCl3)、氘代甲醇(CD3OD)，内标为四甲基硅烷(TMS)。
MS的测定用Agilent 1200/1290 DAD-6110/6120 Quadrupole MS液质联用仪(生产商：Agilent，MS型号：6110/6120 Quadrupole MS)。
waters ACQuity UPLC-QD/SQD(生产商：waters，MS型号：waters ACQuity Qda Detector/waters SQ Detector)
THERMO Ultimate 3000-Q Exactive(生产商：THERMO，MS型号：THERMO Q Exactive)
高效液相色谱法(HPLC)分析使用Agilent HPLC 1200DAD、Agilent HPLC 1200VWD和Waters HPLC e2695-2489高压液相色谱仪。
手性HPLC分析测定使用Agilent 1260 DAD高效液相色谱仪。
高效液相制备使用Waters 2545-2767、Waters 2767-SQ Detecor2、Shimadzu LC-20AP和Gilson GX-281制备型色谱仪。
手性制备使用Shimadzu LC-20AP制备型色谱仪。
CombiFlash快速制备仪使用Combiflash Rf200(TELEDYNE ISCO)。
薄层层析硅胶板使用烟台黄海HSGF254或青岛GF254硅胶板，薄层色谱法(TLC)使用的硅胶板采用的规格是0.15mm～0.2mm，薄层层析分离纯化产品采用的规格是0.4mm～0.5mm。
硅胶柱色谱法一般使用烟台黄海硅胶200～300目硅胶为载体。
激酶平均抑制率及IC50值的测定用NovoStar酶标仪(德国BMG公司)。
本发明的已知的起始原料可以采用或按照本领域已知的方法来合成，或可购买自ABCR GmbH&Co.KG，Acros Organics，Aldrich Chemical Company，韶远化学科技(Accela ChemBio Inc)、达瑞化学品等公司。
实施例中无特殊说明，反应均能够在氩气氛或氮气氛下进行。
氩气氛或氮气氛是指反应瓶连接一个约1L容积的氩气或氮气气球。
氢气氛是指反应瓶连接一个约1L容积的氢气气球。
加压氢化反应使用Parr 3916EKX型氢化仪和清蓝QL-500型氢气发生器或HC2-SS型氢化仪。
氢化反应通常抽真空，充入氢气，反复操作3次。
微波反应使用CEM Discover-S 908860型微波反应器。
实施例中无特殊说明，溶液是指水溶液。
实施例中无特殊说明，反应的温度为室温，为20℃～30℃。
实施例中的反应进程的监测采用薄层色谱法(TLC)，反应所使用的展开剂，纯化化合物采用的柱层析的洗脱剂的体系和薄层色谱法的展开剂体系包括：A：二氯甲烷/甲醇体系，B：正己烷/乙酸乙酯，溶剂的体积比根据化合物的极性不同而进行调节，也可以加入少量的三乙胺和醋酸等碱性或酸性试剂进行调节。
实施例中的化合物含有两个或更多个手性中心时，这些化合物的相对立体化学是通过NMR研究和/或X-射线衍射鉴定的。在这些情况中，使用前缀“rel”、随后使用R/S命名法鉴定这些化合物，此时的R/S仅提供相对立体化学信息，不表示绝对立体化学。例如，表示的1:1混合物，即外消旋体。
实施例1
(±)-rel-(1R,2R,5S)-2-(羟甲基)-3,8-二氮杂双环[3.2.1]辛烷-8-羧酸叔丁酯1g
第一步
(±)-5-甲氧基-3,4-二氢-2H-吡咯-2-羧酸甲酯1b
将(±)-2-吡咯烷酮-5-甲酸甲酯1a(100g，698.61mmol，上海毕得)，硫酸二甲酯(110g，872.10mmol)混合，60℃反应16小时，反应液冷却至室温，冰浴下倒入三乙胺(100g)和甲基叔丁基醚(150mL)的溶液中，用甲基叔丁基醚(300mL×6) 萃取后,减压浓缩即得到粗品标题化合物1b(90g，产率：81.9％)，产品不经纯化直接用于下步反应。
MS m/z(ESI):158.1[M+1]。
第二步
(±)-5-(2-甲氧基-1-硝基-2-氧代乙亚基)吡咯烷-2-羧酸甲酯1c
将粗品化合物1b(90g，572.64mmol)，硝基乙酸甲酯(68.18g，572.63mmol)混合，加热至60℃搅拌反应30小时，反应液冷却至室温后，加入乙酸乙酯(300mL)，搅拌0.5小时后过滤，滤饼干燥后即得到标题化合物1c(70g，产率：50％)，产品不经纯化直接用于下步反应。
MS m/z(ESI):245.1[M+1]。
第三步
4-氧代-3,8-二氮杂双环[3.2.1]辛烷-2-羧酸甲酯(非对映体混合物)1d
将粗品化合物1c(14g，57.3mmol)溶于600mL甲醇中，加入10％钯炭催化剂(湿)(14g)，氢气置换三次，搅拌反应48小时。反应液通过硅藻土过滤，将滤液浓缩即得到粗品标题化合物1d(10g，产率：94.6％)，产品不经纯化直接用于下步反应。
MS m/z(ESI):185.2[M+1]。
第四步
8-(叔丁基)2-甲基(±)-rel-(1R,2R,5S)-4-氧代-3,8-二氮杂双环[3.2.1]辛烷-2,8-二甲酸酯1e
将粗品化合物1d(10g，54.2mmol)溶于300mL二氯甲烷中，冰浴下加入三乙胺(16g，158.12mmol)，二碳酸二叔丁酯(11g，50.4mmol，上海韶远)，搅拌反应14小时，反应液减压浓缩，残余物用硅胶柱色谱法以洗脱剂体系B纯化得到标题化合物1e(3.3g，产率：21.3％)。
MS m/z(ESI):285.2[M+1]。
HPLC分析:保留时间1.02分钟，纯度：98.5％(色谱柱:ACQUITYC18,1.7μm,2.1*50mm；流动相：水(10mM碳酸氢铵)，乙腈，梯度配比：乙腈10％-95％)。
第五步
8-(叔丁基)2-甲基(±)-rel-(1R,2R,5S)-3,8-二氮杂双环[3.2.1]辛烷-2,8-二甲酸酯1f
将化合物1e(400mg，1.4mmol)溶于2mL四氢呋喃中，加入3.5mL 2M的硼烷二甲硫醚络合物四氢呋喃溶液，搅拌反应14小时，反应液加入甲醇淬灭，继续50℃反应14小时，反应液减压浓缩，残余物用硅胶柱色谱法以洗脱剂体系A纯化得到标题化合物1f(176mg，产率：46.2％)。
MS m/z(ESI):271.2[M+1]。
第六步
(±)-rel-(1R,2R,5S)-2-(羟甲基)-3,8-二氮杂双环[3.2.1]辛烷-8-羧酸叔丁酯1g
将化合物1f(1g，3.69μmol)溶于15mL四氢呋喃中，加入4.4mL 1M的氢化铝锂的四氢呋喃溶液，0℃搅拌反应1小时，反应液依次加入0.2mL水，0.2mL15％氢氧化钠水溶液，0.4mL水，再加入无水硫酸钠搅拌10分钟，过滤，将滤液浓缩即得到标题化合物1g(430mg，产率：47.9％)，产品不经纯化直接用于下步反应。
MS m/z(ESI):243.1[M+1]。
实施例2
2,6-二氯-3-氟吡啶-4-胺2a
将化合物1h(5g，30.6mmol)溶于20mL N,N-二甲基甲酰胺和20mL乙腈中，加入1-氯甲基-4-氟-1,4-二氮杂双环[2.2.2]辛烷二(四氟硼酸)盐(13g，36.8mmol)，80℃反应0.5小时，反应液减压浓缩，残余物用硅胶柱色谱法以洗脱剂体系B纯化得到标题化合物3a(2.2g，产率：39.6％)。
MS m/z(ESI):180.9[M+1]。
实施例3
5-乙基-4-((5aS,6S,9R)-1-氟-12-(((2R,7aS)-2-氟四氢-1H-吡咯嗪-7a(5H)-基)甲氧基)-5a,6,7,8,9,10-六氢-5H-4-氧杂-3,10a,11,13,14-五氮杂-6,9-甲桥萘并[1,8-ab]并庚环-2-基)萘-2-酚3-p1
5-乙基-4-((5aR,6R,9S)-1-氟-12-(((2R,7aS)-2-氟四氢-1H-吡咯嗪-7a(5H)-基)甲氧基)-5a,6,7,8,9,10-六氢-5H-4-氧杂-3,10a,11,13,14-五氮杂-6,9-甲桥萘并[1,8-ab]并庚环-2-基)萘-2-酚3-p2
第一步
(±)-rel-(1R,2R,5S)-2-(((叔丁基二甲基硅)氧基)甲基)-3,8-二氮杂双环[3.2.1]辛烷-8-羧酸叔丁酯3a
将化合物1g(8.8g，36.3mmol)，叔丁基二甲基氯硅烷(16g,106.1558mmol)，4-二甲氨基吡啶(4g,32.4739mmol)溶于200mL二氯甲烷中，加入三乙胺(15g,148.23mmol,21.4286mL)，搅拌反应16小时，反应液减压浓缩，残余物用硅胶柱色谱法以洗脱剂体系B纯化得到标题化合物3a(8g，产率：61.7％)。
MS m/z(ESI):357.1[M+1]。
第二步
4-((叔丁氧基羰基)氨基)-2,6-二氯-5-氟烟酸叔丁酯3b
将化合物2a(1.8g，9.94mmol)溶于四氢呋喃(50mL)，冰浴下加入20mL 2M的双三甲基硅基氨基钠四氢呋喃溶液，搅拌反应0.5小时后，加入二碳酸二叔丁酯(6.5g，29.7mmol)，搅拌反应14小时，反应液中加入饱和氯化铵水溶液淬灭，乙酸乙酯萃取(50mL×3)，有机相合并，用无水硫酸钠干燥，过滤除去干燥剂后滤液减压浓缩，残余物以洗脱剂体系B纯化得到标题化合物3b(1g，产率：26.3％)，产品不经纯化直接用于下步反应。
MS m/z(ESI):381.1[M+1]。
第三步
4-氨基-2,6-二氯-5-氟烟酸叔丁酯3c
将化合物3b(1g，2.62mmol)溶于乙酸乙酯(8mL)中，加入3mL 4M的盐酸二氧六环溶液，搅拌反应2小时，冰浴下用4M的氢氧化钠水溶液调节pH至中性，乙酸乙酯萃取(10mL×3)，有机相合并，用无水硫酸钠干燥，过滤除去干燥剂后滤液减压浓缩，残余物以洗脱剂体系B纯化得到粗品标题化合物3c(500mg，产率：67.8％)。
MS m/z(ESI):281.1[M+1]。
第四步
2,6-二氯-5-氟-4-(3-(2,2,2-三氯乙酰基)脲基)烟酸叔丁酯3d
将粗品化合物3c(500mg，1.77mmol)溶于四氢呋喃(10mL)，加入三氯乙酰异氰酸酯(670mg，3.55mmol)，搅拌反应30分钟，反应液减压浓缩即得到粗品标题化合物3d(835mg，产率：99.7％)，产品不经纯化直接用于下步反应。
MS m/z(ESI):467.9[M+1]。
第五步
5,7-二氯-8-氟-吡啶并[4,3-d]嘧啶-2,4-二酚3e
将粗品化合物3d(835mg，1.77mmol)溶于7M的氨甲醇溶液(10mL)，搅拌反应1小时，反应液减压浓缩，残余物中加入甲基叔丁基醚(10mL)，搅拌0.5小时后过滤，滤饼干燥后即得到粗品标题化合物3e(400mg，产率：89.9％)，产品不经纯化直接用于下步反应。
MS m/z(ESI):249.9[M+1]。
第六步
2,4,5,7-四氯-8-氟-吡啶并[4,3-d]嘧啶3f
将粗品化合物3e(300mg，1.19mmol)溶于三氯氧磷(6mL)，加入N,N-二异丙基乙胺(800mg，6.19mmol)，110℃搅拌反应3小时，反应液冷却至室温后减压浓缩，得粗品标题化合物3f(344mg，产率：97.7％)，产物不经纯化直接用于下一步。
MS m/z(ESI):285.8[M+1]。
第七步
(±)-rel-(1R,2R,5S)-2-(((叔丁基二甲基硅基)氧基)甲基)-3-(2,5,7-三氯-8-氟吡啶并[4,3-d]嘧啶-4-基)-3,8-二氮杂双环[3.2.1]辛烷-8-羧酸叔丁酯3g
将化合物3f(1.0g，3.48mmol)，N,N-二异丙基乙胺(0.9g，6.9mmol)溶于15mL二氯甲烷中，-78℃加入3a(1.25g，3.5mmol)，保持温度搅拌反应1小时后恢复室温反应16小时，反应液减压浓缩，残余物以洗脱剂体系B纯化得到粗品标题化合物3g(1.56g，产率：73.7％)。
MS m/z(ESI):606.2[M+1]。
第八步
(1S,2S,5R)-2-(((叔丁基二甲基硅基)氧基)甲基)-3-(5,7-二氯-8-氟-2-(((2R,7aS)-2-氟四氢-1H-吡咯嗪-7a(5H)-基)甲氧基)吡啶并[4,3-d]嘧啶-4-基)-3,8-二氮杂双环[3.2.1]辛烷-8-羧酸叔丁酯3h-1
和
(1R,2R,5S)-2-(((叔丁基二甲基硅基)氧基)甲基)-3-(5,7-二氯-8-氟-2-(((2R,7aS)-2-氟四氢-1H-吡咯嗪-7a(5H)-基)甲氧基)吡啶并[4,3-d]嘧啶-4-基)-3,8-二氮杂双环[3.2.1]辛烷-8-羧酸叔丁酯3h-2的非对映体混合物
将化合物3g(1.4g，2.3mmol)溶于1,4-二氧六环(20mL)，加入((2R,7aS)-2-氟四氢-1H-吡咯嗪-7a(5H)-基)甲醇(650mg，4.08mmol，药明)，N,N-二异丙基乙胺(1.5g，11.6mmol)，4A分子筛(1.4g)，105℃搅拌反应6小时，反应液冷却至室温后过滤，减压浓缩，得粗品标题化合物3h-1和3h-2的非对映体混合物(1.68g，产率：99.8％)，产物不经纯化直接用于下一步。
MS m/z(ESI):729.2[M+1]
第九步
(5aS,6S,9R)-2-氯-1-氟-12-(((2R,7aS)-2-氟四氢-1H-吡咯嗪-7a(5H)-基)甲氧基)-5a,6,7,8,9,10-六氢-5H-4-氧杂-3,10a,11,13,14-五氮杂-6,9-甲桥萘并[1,8-ab]并庚环-14-羧酸叔丁酯3i-1
和
(5aR,6R,9S)-2-氯-1-氟-12-(((2R,7aS)-2-氟四氢-1H-吡咯嗪-7a(5H)-基)甲氧基)-5a,6,7,8,9,10-六氢-5H-4-氧杂-3,10a,11,13,14-五氮杂-6,9-甲桥萘并[1,8-ab]并庚环-14-羧酸叔丁酯3i-2的非对映体混合物
将粗品化合物3h-1和3h-2的非对映体混合物(1.68g，2.3mmol)，加入四丁基氟化铵(2.59g，11.51mmol)，室温搅拌16小时；反应液减压浓缩，残余物以洗脱剂体系B纯化得到标题化合物3i-1和3i-2的非对映体混合物(1.0g，产率：75.0％)。
MS m/z(ESI):579.2[M+1]。
第十步
(5aS,6S,9R)-2-(8-乙基-3-(甲氧基甲氧基)萘-1-基)-1-氟-12-(((2R,7aS)-2-氟四氢-1H-吡咯嗪-7a(5H)-基)甲氧基)-5a,6,7,8,9,10-六氢-5H-4-氧杂-3,10a,11,13,14-五氮杂-6,9-甲桥萘并[1,8-ab]并庚环-14-羧酸叔丁基酯3j-1
和
(5aR,6R,9S)-2-(8-乙基-3-(甲氧基甲氧基)萘-1-基)-1-氟-12-(((2R,7aS)-2-氟四氢-1H-吡咯嗪-7a(5H)-基)甲氧基)-5a,6,7,8,9,10-六氢-5H-4-氧杂-3,10a,11,13,14-五氮杂-6,9-甲桥萘并[1,8-ab]并庚环-14-羧酸叔丁基酯3j-2的非对映体1:1混合物
将化合物3i-1和3i-2的非对映体混合物(1.9g，3.28mmol)，2-(8-乙基-3-(甲氧基甲氧基)萘-1-基)-4,4,5,5-四甲基-1,3,2-二氧硼杂环戊烷(1.5g，4.38mmol，采用专利申请“WO2021/041671”中说明书第111页的中间体21公开的方法制备而得)，四(三苯基膦)钯(1.16g，1mmol，adamas)，碳酸铯(4.7g，14.4mmol)溶于36mL 1,4-二氧六环和水(V:V＝5:1)的混合溶液中。氮气氛围下，100℃反应6小时，反应液减压浓缩即得粗品标题化合物3j-1和3j-2的非对映体1:1混合物(2.5g，产率：100％)。
MS m/z(ESI):759.2[M+1]。
第十一步
5-乙基-4-((5aS,6S,9R)-1-氟-12-(((2R,7aS)-2-氟四氢-1H-吡咯嗪-7a(5H)-基)甲氧基)-5a,6,7,8,9,10-六氢-5H-4-氧杂-3,10a,11,13,14-五氮杂-6,9-甲桥萘并[1,8-ab]并庚环-2-基)萘-2-酚3p-1
5-乙基-4-((5aR,6R,9S)-1-氟-12-(((2R,7aS)-2-氟四氢-1H-吡咯嗪-7a(5H)-基)甲氧基)-5a,6,7,8,9,10-六氢-5H-4-氧杂-3,10a,11,13,14-五氮杂-6,9-甲桥萘并[1,8-ab]并庚环-2-基)萘-2-酚3p-2
将粗品化合物3j-1和3j-2的非对映体混合物(2.4g，3.16mol)溶于乙酸乙酯(40mL)中，加入17mL 4M的盐酸二氧六环溶液，0℃反应2小时，反应液减压浓缩，残余物用高效液相色谱法(Waters-2545，色谱柱：SharpSil-T C18,30*150mm,5μm；流动相：水相(10mmol/L碳酸氢铵)和乙腈，梯度配比：乙腈38％-45％，流速：30mL/min)纯化得到标题化合物3p-1和3p-2的非对映体1:1混合物(620mg，产率：32.6％)。
MS m/z(ESI):615.2[M+1]。
1H NMR(500MHz,CD3OD):δ7.63(d,1H),7.37(q,1H),7.29(q,1H),7.18(dd,1H),7.08(t,1H),5.48-5.29(m,2H),4.55-4.28(m,4H),4.17(dd,1H),3.84-3.75(m,1H),3.70(t,1H),3.47(q,1H),3.31-3.07(m,3H),2.57-2.17(m,5H),2.15-1.79(m,7H),1.63(t,1H),1.08-0.87(m,3H)。
化合物3p-1和3p-2的非对映体混合物经手性柱拆分(Shimadzu LC-20AP，色谱柱：DAICEL25*250mm,10μm；流动相A：正己烷，流动相B：乙醇(0.1％7M NH3in MeOH))和，梯度配比：A:B：30:70，流速：30mL/min)得到标题化合物3p-1(50mg，产率：35.7％)和3p-2(50mg，产率：35.7％)。
单一构型化合物(较短保留时间)3p-2：(50mg，产率：35.7％)。
MS m/z(ESI):615.2[M+1]。
手性HPLC分析：保留时间9.85分钟，纯度：99％(色谱柱：DAICEL250*4.6mm，5μm；流动相：正己烷和乙醇(含0.2％二乙胺)，流速：1.0mL/min)。
1H NMR(500MHz,CD3OD):δ7.61-7.59(m,1H),7.36-7.32(m,1H),7.26-7.25(m,1H),7.18-7.13(m 1H),7.05-6.95(m,1H),5.37-5.22(m,2H),5.09-5.00(m,1H),4.61-4.56(m,1H),4.49-4.41(m,1H),4.30(dd,1H),4.24-4.18(m,1H),4.16-4.09(m,1H),3.72(dd,1H),3.62(dd,1H),3.27-3.17(m,3H),3.01(td,1H),2.48(dt,1H),2.40–2.10(m,5H),1.99(td,2H),1.94-1.75(m,4H),0.99-0.88m,3H)。
单一构型化合物(较长保留时间)3p-1：(50mg，产率：35.7％)。
MS m/z(ESI):615.2[M+1]。
手性HPLC分析：保留时间16.0分钟，纯度：99％(色谱柱：DAICEL250*4.6mm，5μm；流动相：正己烷和乙醇(含0.2％二乙胺)，流速：1.0mL/min)。
1H NMR(500MHz,CD3OD):δ7.61-7.59(m,1H),7.36-7.32(m,1H),7.26-7.25(m,1H),7.18-7.13(m 1H),7.05-6.94(m,1H),5.36-5.33(m,2H),5.10-5.01(m,1H),4.61-4.56(m,1H),4.49-4.41(m,1H),4.30(dd,1H),4.24-4.22(m,1H),4.16-4.10(m,1H),3.73-3.72(m,1H),3.64-3.61(m,1H),3.26-3.20(m,3H),3.04-2.99(m,1H),2.48(dt,1H),2.38-2.17(m,5H),2.03-1.96(m,2H),1.93-1.78(m,4H),0.99-0.88m,3H)。
实施例4
5-乙基-6-氟-4-((5aS,6S,9R)-1-氟-12-(((2R,7aS)-2-氟四氢-1H-吡咯嗪-7a(5H)-基)甲氧基)-5a,6,7,8,9,10-六氢-5H-4-氧杂-3,10a,11,13,14-五氮杂-6,9-甲桥萘并[1,8-ab]并庚环-2-基)萘-2-酚4p-1
5-乙基-6-氟-4-((5aR,6R,9S)-1-氟-12-(((2R,7aS)-2-氟四氢-1H-吡咯嗪-7a(5H)-基)甲氧基)-5a,6,7,8,9,10-六氢-5H-4-氧杂-3,10a,11,13,14-五氮杂-6,9-甲桥萘并[1,8-ab]并庚环-2-基)萘-2-酚4p-2
第十步
(5aS,6S,9R)-2-(8-乙基-7-氟-3-(甲氧基甲氧基)萘-1-基)-1-氟-12-(((2R,7aS)-2-氟四氢-1H-吡咯嗪-7a(5H)-基)甲氧基)-5a,6,7,8,9,10-六氢-5H-4-氧杂-3,10a,11,13,14-五氮杂-6,9-甲桥萘并[1,8-ab]并庚环-14-羧酸叔丁酯4a-1
和
(5aR,6R,9S)-2-(8-乙基-7-氟-3-(甲氧基甲氧基)萘-1-基)-1-氟-12-(((2R,7aS)-2-氟四氢-1H-吡咯嗪-7a(5H)-基)甲氧基)-5a,6,7,8,9,10-六氢-5H-4-氧杂-3,10a,11,13,14-五氮杂-6,9-甲桥萘并[1,8-ab]并庚环-14-羧酸叔丁酯4a-2的非对映体1:1混合物
将化合物3i-1和3i-2的非对映体混合物(300mg，518.1μmol)，2-(8-乙基-7-氟-3-(甲氧基甲氧基)萘-1-基)-4,4,5,5-四甲基-1,3,2-二氧硼杂环戊烷(280mg，777.2μmol，采用专利申请“WO2021/041671”中说明书第104页的中间体18公开的方法制备而得)，四(三苯基膦)钯(120mg，103.8μmol)，碳酸铯(506mg，1.55mmol)溶于6mL 1,4-二氧六环和水(V:V＝5:1)的混合溶液中。氮气氛围下，100℃反应6小时，反应液减压浓缩即得粗品标题化合物4a-1和4a-2的非对映体1:1混合物(400mg)，产物不经纯化，直接用于下步反应。
MS m/z(ESI):777.2[M+1]。
第十一步
5-乙基-6-氟-4-((5aS,6S,9R)-1-氟-12-(((2R,7aS)-2-氟四氢-1H-吡咯嗪-7a(5H)-基)甲氧基)-5a,6,7,8,9,10-六氢-5H-4-氧杂-3,10a,11,13,14-五氮杂-6,9-甲桥萘并[1,8-ab]并庚环-2-基)萘-2-酚4p-1
5-乙基-6-氟-4-((5aR,6R,9S)-1-氟-12-(((2R,7aS)-2-氟四氢-1H-吡咯嗪-7a(5H)-基)甲氧基)-5a,6,7,8,9,10-六氢-5H-4-氧杂-3,10a,11,13,14-五氮杂-6,9-甲桥萘并[1,8-ab]并庚环-2-基)萘-2-酚4p-2
将粗品化合物4a-1和4a-2的非对映体混合物(160mg，205.9μmol)溶于乙酸乙酯(5mL)中，加入1mL 4M的盐酸二氧六环溶液，0℃反应2小时，反应液减压浓缩，残余物用高效液相色谱法(Waters-2545，色谱柱：SharpSil-T C18,30*150mm,5μm；流动相：水相(10mmol/L碳酸氢铵)和乙腈，梯度配比：乙腈38％-45％，流速：30mL/min)纯化得到标题化合物4p-1(即式I所示化合物)和4p-2的非对映体1:1混合物(10mg，产率：7.2％)。
MS m/z(ESI):633.2[M+1]。
1H NMR(500MHz,CD3OD):δ7.67(ddd,1H),7.32-7.21(m,2H),7.11-7.01(m,1H),5.38-5.35(m,2H),5.11-5.03(m,1H),4.64-4.59(m,1H),4.52-4.46(m,1H),4.34-4.29(m,1H),4.25(dd,1H),4.18-4.12(m,1H),3.74(br,1H),3.65(br,1H),3.26-3.23(m,3H),3.05-3.01(m,1H),2.61-1.81(m,12H),0.94-0.82(m,3H)。
化合物4p-1和4p-2的非对映体混合物经手性柱拆分(Shimadzu LC-20AP，色谱柱：DAICEL25*250mm,10μm；流动相A：正己烷，流动相B：乙醇(0.1％7M NH3in MeOH))和，梯度配比：A:B：40:60，流速：30mL/min)得到标题化合物4p-1(26mg，产率：43.3％)和4p-2(26mg，产率：43.3％)。
单一构型化合物(较短保留时间)4p-2：(26mg，产率：43.3％)。
MS m/z(ESI):633.2[M+1]。
手性HPLC分析：保留时间7.89分钟，纯度：99％(色谱柱：DAICEL250*4.6mm，5μm；流动相：正己烷和乙醇(含0.2％二乙胺)，流速：1.0mL/min)。
1H NMR(500MHz,CD3OD):δ7.67(ddd,1H),7.32-7.21(m,2H),7.11-7.01(m,1H),5.38-5.27(m,2H),5.11-5.03(m,1H),4.64-4.59(m,1H),4.52-4.44(m,1H),4.33(d,1H),4.24(dd,1H),4.18-4.12(m,1H),3.75(br,1H),3.66(br,1H),3.27-3.18(m,3H),3.05-3.03(m,1H),2.60-1.81(m,12H),0.93-0.82(m,3H)。
单一构型化合物(较长保留时间)4p-1：(26mg，产率：43.3％)。
MS m/z(ESI):633.2[M+1]。
手性HPLC分析：保留时间13.8分钟，纯度：99％(色谱柱：DAICEL250*4.6mm，5μm；流动相：正己烷和乙醇(含0.2％二乙胺)，流速：1.0mL/min)。
1H NMR(500MHz,CD3OD):δ7.67(ddd,1H),7.32-7.21(m,2H),7.11-7.01(m,1H),5.38-5.35(m,2H),5.11-5.03(m,1H),4.64-4.59(m,1H),4.52-4.46(m,1H),4.34-4.29(m,1H),4.25(dd,1H),4.18-4.12(m,1H),3.74(br,1H),3.65(br,1H),3.26-3.23(m,3H),3.05-3.01(m,1H),2.61-1.81(m,12H),0.94-0.82m,3H)。
实施例5
称取葡萄糖酸铜，加适量的水搅拌溶解均匀，用三乙醇胺将pH值调节至7.4，定容至180ml，得到水相溶液1。称取盐酸组氨酸和氯化钠，以适量水溶解后，用氢氧化钠和/或盐酸调节pH至7.4，定容至2400ml，得到水相溶液2。
称取氢化大豆磷脂酰胆碱、胆固醇、培化磷脂酰乙醇胺，加入无水乙醇后加热搅拌溶解，得到脂质溶液。将脂质溶液加入上述水相溶液1中，在高于磷脂相变温度的条件下搅拌约15分钟，之后继续水化并减压去除绝大部分乙醇，通过挤出器挤出整粒得到空白脂质体。以切向流过滤系统对空白脂质体进行处理，将脂质体外水相替换为水相溶液2。将式I化合物溶解于乙醇溶液中，加入适量的盐酸，得到120mg/ml的药物溶液。将药物溶液缓慢加入空白脂质体中，在高于磷脂相变温度的条件下孵育15分钟后，迅速降温至室温，以切向流系统将产品浓缩至目标浓度并进一步除去残留乙醇，定容，得到脂质体产品，产品表征信息如下表。

实施例6
称取硫酸铵、磷酸，加适量的水搅拌溶解均匀，用氨水将pH值调节至5.85，定容至180ml，得到水相溶液1。称取盐酸组氨酸和氯化钠，以适量水溶解后，用氢氧化钠和/或盐酸调节pH至6.35，定容至2400ml，得到水相溶液2。
称取氢化大豆磷脂酰胆碱、胆固醇、培化磷脂酰乙醇胺，加入无水乙醇后加热搅拌溶解，得到脂质溶液。将脂质溶液加入上述水相溶液1中，在高于磷脂相变温度的条件下搅拌约15分钟，之后继续水化并减压去除绝大部分乙醇，通过挤出器挤出整粒得到空白脂质体。以切向流过滤系统对空白脂质体进行处理，将脂质体外水相替换为水相溶液2。将式I化合物溶解于乙醇溶液中，加入适量的盐酸，得到120mg/ml的药物溶液。将药物溶液缓慢加入空白脂质体中，在高于磷脂相变温度的条件下孵育15分钟后，迅速降温至室温，以切向流系统将产品浓缩至目标浓度并进一步除去残留乙醇，定容，得到脂质体产品，产品表征信息如下表。
实施例7
称取硫酸铵、磷酸，加适量的水搅拌溶解均匀，用氨水将pH值调节至7.0，定容至180ml，得到水相溶液1。称取盐酸组氨酸和氯化钠，以适量水溶解后，用氢氧化钠和/或盐酸调节pH至7.5，定容至2400ml，得到水相溶液2。
称取氢化大豆磷脂酰胆碱、胆固醇、培化磷脂酰乙醇胺，加入无水乙醇后加热搅拌溶解，得到脂质溶液。将脂质溶液加入上述水相溶液1中，在高于磷脂相变温度的条件下搅拌约15分钟，之后继续水化并减压去除绝大部分乙醇，通过挤出器挤出整粒得到空白脂质体。以切向流过滤系统对空白脂质体进行处理，将脂质体外水相替换为水相溶液2。将式I化合物溶解于乙醇溶液中，加入适量的盐酸，得到120mg/ml的药物溶液。将药物溶液缓慢加入空白脂质体中，在高于磷脂相变温度的条件下孵育15分钟后，迅速降温至室温，以切向流系统将产品 浓缩至目标浓度并进一步除去残留乙醇，定容，得到脂质体产品，产品表征信息如下表。
实施例8
称取硫酸铵、柠檬酸一水合物，加适量的水搅拌溶解均匀，用氨水将pH值调节至7.0，定容至180ml，得到水相溶液1。称取盐酸组氨酸和氯化钠，以适量水溶解后，用氢氧化钠和/或盐酸调节pH至7.5，定容至2400ml，得到水相溶液2。
称取氢化大豆磷脂酰胆碱、胆固醇、培化磷脂酰乙醇胺，加入无水乙醇后加热搅拌溶解，得到脂质溶液。将脂质溶液加入上述水相溶液1中，在高于磷脂相变温度的条件下搅拌约15分钟，之后继续水化并减压去除绝大部分乙醇，通过挤出器挤出整粒得到空白脂质体。以切向流过滤系统对空白脂质体进行处理，将脂质体外水相替换为水相溶液2。将式I化合物溶解于乙醇溶液中，加入适量的盐酸，得到120mg/ml的药物溶液。将药物溶液缓慢加入空白脂质体中，在高于磷脂相变温度的条件下孵育15分钟后，迅速降温至室温，以切向流系统将产品 浓缩至目标浓度并进一步除去残留乙醇，定容，得到脂质体产品，产品表征信息如下表。
实施例9
称取硫酸铵、苹果酸，加适量的水搅拌溶解均匀，用氨水将pH值调节至7.0，定容至180ml，得到水相溶液1。称取盐酸组氨酸和氯化钠，以适量水溶解后，用氢氧化钠和/或盐酸调节pH至7.5，定容至2400ml，得到水相溶液2。
称取氢化大豆磷脂酰胆碱、胆固醇、培化磷脂酰乙醇胺，加入无水乙醇后加热搅拌溶解，得到脂质溶液。将脂质溶液加入上述水相溶液1中，在高于磷脂相变温度的条件下搅拌约15分钟，之后继续水化并减压去除绝大部分乙醇，通过挤出器挤出整粒得到空白脂质体。以切向流过滤系统对空白脂质体进行处理，将脂质体外水相替换为水相溶液2。将式I化合物溶解于乙醇溶液中，加入适量的盐酸，得到120mg/ml的药物溶液。将药物溶液缓慢加入空白脂质体中，在高于磷脂相变温度的条件下孵育15分钟后，迅速降温至室温，以切向流系统将产品 浓缩至目标浓度并进一步除去残留乙醇，定容，得到脂质体产品，产品表征信息如下表。
实施例10
称取硫酸铵、盐酸组氨酸，加适量的水搅拌溶解均匀，用氨水将pH值调节至7.0，定容至180ml，得到水相溶液1。称取盐酸组氨酸和氯化钠，以适量水溶解后，用氢氧化钠和/或盐酸调节pH至7.5，定容至2400ml，得到水相溶液2。
称取氢化大豆磷脂酰胆碱、胆固醇、培化磷脂酰乙醇胺，加入无水乙醇后加热搅拌溶解，得到脂质溶液。将脂质溶液加入上述水相溶液1中，在高于磷脂相变温度的条件下搅拌约15分钟，之后继续水化并减压去除绝大部分乙醇，通过挤出器挤出整粒得到空白脂质体。以切向流过滤系统对空白脂质体进行处理，将脂质体外水相替换为水相溶液2。将式I化合物溶解于乙醇溶液中，加入适量的盐酸，得到120mg/ml的药物溶液。将药物溶液缓慢加入空白脂质体中，在高于磷脂相变温度的条件下孵育15分钟后，迅速降温至室温，以切向流系统将产品 浓缩至目标浓度并进一步除去残留乙醇，定容，得到脂质体产品，产品表征信息如下表。
实施例11
称取硫酸铵、磷酸，加适量的水搅拌溶解均匀，用氨水将pH值调节至5.85，定容至180ml，得到水相溶液1。称取盐酸组氨酸和氯化钠，以适量水溶解后，用氢氧化钠和/或盐酸调节pH至6.35，定容至2400ml，得到水相溶液2。
称取氢化大豆磷脂酰胆碱、胆固醇、培化磷脂酰乙醇胺，加入无水乙醇后加热搅拌溶解，得到脂质溶液。将脂质溶液加入上述水相溶液1中，在高于磷脂相变温度的条件下搅拌约15分钟，之后继续水化并减压去除绝大部分乙醇，通过挤出器挤出整粒得到空白脂质体。以切向流过滤系统对空白脂质体进行处理，将脂质体外水相替换为水相溶液2。将式I化合物溶解于乙醇溶液中，加入适量的盐酸，得到120mg/ml的药物溶液。将药物溶液缓慢加入空白脂质体中，在高于磷脂相变温度的条件下孵育15分钟后，迅速降温至室温，以切向流系统将产品 浓缩至目标浓度并进一步除去残留乙醇，定容，得到脂质体产品，产品表征信息如下表。
实施例12
称取硫酸铵、磷酸，加适量的水搅拌溶解均匀，用氨水将pH值调节至5.85，定容至180ml，得到水相溶液1。称取盐酸组氨酸和氯化钠，以适量水溶解后，用氢氧化钠和/或盐酸调节pH至6.35，定容至2400ml，得到水相溶液2。
称取氢化大豆磷脂酰胆碱、胆固醇、培化磷脂酰乙醇胺，加入无水乙醇后加热搅拌溶解，得到脂质溶液。将脂质溶液加入上述水相溶液1中，在高于磷脂相变温度的条件下搅拌约15分钟，之后继续水化并减压去除绝大部分乙醇，通过挤出器挤出整粒得到空白脂质体。以切向流过滤系统对空白脂质体进行处理，将脂质体外水相替换为水相溶液2。将式I化合物溶解于乙醇溶液中，加入适量的盐酸，得到120mg/ml的药物溶液。将药物溶液缓慢加入空白脂质体中，在高于磷脂相变温度的条件下孵育15分钟后，迅速降温至室温，以切向流系统将产品 浓缩至目标浓度并进一步除去残留乙醇，定容，得到脂质体产品，产品表征信息如下表。
生物学评价
测试例1：AGS细胞ERK磷酸化抑制实验生物学评价(HTRF法)
一、测试目的
本实验通过检测式I所示化合物对细胞ERK磷酸化抑制作用，根据IC50大小评价化合物对KRAS靶点的抑制作用。
二、实验方法
AGS细胞(南京科佰，CBP60476)用含有10％胎牛血清的RPMI1640(Hyclone，SH30809.01)完全培养基进行培养。实验第一天，使用完全培养基将AGS细胞以40000个/孔的密度种于96孔板，每孔190μL细胞悬液，放置37℃，5％CO2细胞培养箱培养过夜。
第二天，每孔加入10μL用完全培养基配制的梯度稀释的待测化合物，化合物的终浓度是从10μM开始进行5倍梯度稀释的9个浓度点，设置含有0.5％DMSO的空白对照，孔板放置37℃，5％CO2的细胞培养箱孵育1个小时。孵育完成后，取出96孔细胞培养板，吸掉培养基，每孔加入200μL PBS(上海源培生物科技股份有限公司，B320)洗一遍。吸掉PBS，每孔加入50μL含封闭液(blocking reagent，Cisbio，64KB1AAC)的裂解缓冲液(lysis buffer，Cisbio，64KL1FDF)，孔板放置振荡器上室温震荡裂解40分钟。裂解后用移液器吹打混匀，每孔各转移16μL裂解液分别至两块HTRF 96孔检测板(Cisbio，66PL96100)中，之后两块板分别加入4μL预混的磷酸化ERK1/2抗体溶液(Cisbio，64AERPEG)或4μL预混的总ERK1/2抗体溶液(Cisbio，64NRKPEG)。微孔板用封板膜密封，在微孔板离心机中离心1分钟，室温避光孵育过夜。
第三天，使用ENVISION多功能酶标仪(PerkinElmer，ENVISION)读取337nm波长激发，665nm和620nm波长发射的荧光值。
三、数据分析与结果
用Graphpad Prism软件根据化合物浓度和磷酸化ERK/总ERK的比值计算化合物抑制活性的IC50值。结果参见下表1。
表1 AGS细胞ERK磷酸化抑制活性数据
结论：式I所示化合物对AGS细胞ERK磷酸化具有较好的抑制作用。
测试例2：GP2d、AGS细胞3D增殖抑制实验生物学评价
一、测试目的
通过测试式I所示化合物对GP2d、AGS细胞的3D增殖抑制作用，评价式I所示化合物对KRAS靶点的抑制作用。
二、实验方法
GP2d细胞(南京科佰，CBP60010)用完全培养基即含有10％胎牛血清(Corning，35-076-CV)的DMEM/高糖培养基(Hyclone，SH30243.01)进行培养。实验第一天，使用完全培养基将GP2d细胞以1000个细胞/孔的密度种于96孔低吸附板(Corning，CLS7007-24EA)，每孔90μL细胞悬液，2000rpm室温离心5分钟后放置37℃，5％CO2细胞培养箱培养过夜。
AGS细胞(南京科佰，CBP60476)用完全培养基即含有10％胎牛血清(Corning，35-076-CV)的RPMI1640培养基(Hyclone，SH30809.01)进行培养。实验第一天，使用完全培养基将AGS细胞以1000个细胞/孔的密度种于96孔低吸附板(Corning，CLS7007-24EA)，每孔90μL细胞悬液，2000rpm室温离心5分钟后放置37℃，5％CO2细胞培养箱培养过夜。
第二天，每孔加入10μL用完全培养基配制的梯度稀释的待测化合物，GP2d细胞的化合物终浓度是从1μM开始进行3倍梯度稀释的9个浓度点，AGS细胞的化合物终浓度是从10μM开始进行3倍梯度稀释的9个浓度点，都设置含有0.5％DMSO的空白对照。孔板放置37℃，5％CO2的细胞培养箱培养120小时。第七天，取出96孔细胞培养板，每孔加入50μL3D试剂(Promega，G9682)，室温震荡25分钟后，吹吸混匀并取出50μL转移至白色不透底的96孔板(PE，6005290)中，使用多功能微孔板酶标仪(PerkinElmer，ENVISION)读取发光信号值。
三、数据分析与结果
用Graphpad Prism软件计算化合物抑制活性的IC50值，结果参见下表2。
表2 AGS、GP2d细胞3D增殖抑制活性数据
结论：式I所示化合物对AGS、GP2d细胞3D增殖具有较好的抑制作用。
测试例3：SPR方法检测化合物与KRAS蛋白亚型G12D或WT的亲和力
先将生物素化的Avi-KRAS-WT或Avi-KRAS-G12D用含有100mM MgCl2的1×HBS-P+(Cat.#BR1006-71)缓冲液稀释至20μg/mL，然后流过SA(Cat.#BR1005-31)生物传感芯片通道2，持续420s，获得大约5000-7000RU的偶联水平。再将化合物样品从高到低依次进样120s，之后解离720s。试验采用单循环动力学模式。Biacore 8K仪器实时检测反应信号获得结合解离曲线。试验结束后用Biacore 8K评价软件进行数据分析，采用1:1模型进行数据拟合并获得亲和力数据，结果参见下表3。
结论：式I所示化合物与KRAS蛋白亚型G12D或WT具有较好的亲和力。

Claims (16)

  1. 一种药物组合物，其包含式I化合物或其可药用盐和脂质，
  2. 根据权利要求1所述的药物组合物，其中所述的药物组合物为脂质体。
  3. 根据权利要求1或2所述的药物组合物，其中所述脂质包含至少一种磷脂，所述磷脂优选二棕榈酰磷脂酰胆碱(DPPC)、二硬脂酰磷脂酰胆碱(DSPC)、二肉豆蔻酰磷脂酰胆碱(DMPC)、1-棕榈酰-2-亚油酰-sn-甘油-3-磷脂酰胆碱(PLPC)、二油酰磷脂酰胆碱(DOPC)、二芥酰磷脂酰胆碱(DEPC)、蛋黄磷脂酰胆碱(EPC)、二月桂酰磷脂酰胆碱(DLPC)、氢化大豆磷脂酰胆碱(HSPC)、l-肉豆蔻酰-2-棕榈酰磷脂酰胆碱(MPPC)、l-棕榈酰-2-肉豆蔻酰磷脂酰胆碱(PMPC)、l-棕榈酰-2-硬脂酰磷脂酰胆碱(PSPC)、l-硬脂酰-2-棕榈酰磷脂酰胆碱(SPPC)、棕榈酰油酰磷脂酰胆碱(POPC)、溶血磷脂酰胆碱、二亚油酰磷脂酰胆碱、二硬脂酰磷脂酰乙醇胺(DSPE)、二肉豆蔻酰磷脂酰乙醇胺(DMPE)、二棕榈酰磷脂酰乙醇胺(DPPE)、二油酰基磷脂酰甘油(DOPG)、二肉豆蔻酰磷脂酰甘油(DMPG)、二硬脂酰基磷脂酰甘油(DSPG)、二棕榈酰基甘油磷酸甘油(DPPG)、二棕榈酰基磷脂酰丝氨酸(DPPS)、1，2-二油酰基-sn-甘油-3-磷脂酰丝氨酸(DOPS)、二肉豆蔻酰基磷脂酰丝氨酸(DMPS)、二硬脂酰基磷脂酰丝氨酸(DSPS)、二棕榈酰基磷脂酸(DPPA)、1，2-二油酰基-sn-甘油-3-磷脂酸(DOPA)、二肉豆蔻酰基磷脂酸(DMPA)、二硬脂酰基磷脂酸(DSPA)、二棕榈酰基磷脂酰肌醇(DPPI)、1，2-二油酰基-sn-甘油-3-磷脂酰肌醇(DOPI)、二肉豆蔻酰基磷脂酰肌醇(DMPI)、二硬脂酰基磷脂酰肌醇(DSPI)，以及它们的衍生物，更优选二棕榈酰磷脂酰胆碱(DPPC)、二硬脂酰磷脂酰胆碱(DSPC)、二油酰磷脂酰胆碱(DOPC)、二芥酰磷脂酰胆碱(DEPC)、二月桂酰磷脂酰胆碱(DLPC)、氢化大豆磷脂酰胆碱(HSPC)、二硬脂酰磷脂酰乙醇胺(DSPE)和二肉豆蔻酰磷脂酰胆碱(DMPC)以及它们的聚乙二醇修饰的衍生物，最优选氢化大豆磷脂酰胆碱(HSPC)和聚乙二醇2000-二硬脂酰磷脂酰乙醇胺(mPEG2000-DSPE)。
  4. 根据权利要求1-3任意一项所述的药物组合物，相对于总脂质混合物的量，所述磷脂的摩尔百分比选自10％～80％，优选30％～70％。
  5. 根据权利要求1-4任意一项所述的药物组合物，所述的脂质还包含类固醇，优选胆固醇。
  6. 根据权利要求5所述的药物组合物，其中相对于总脂质混合物的量，类固醇的摩尔百分比选自0.1％-90％，优选10％-80％，更优选20％-70％。
  7. 根据权利要求1-6任意一项所述的药物组合物，其中还包含捕获剂，所述捕获剂优选硫酸盐、亚硫酸盐、葡萄糖酸盐、磷酸盐、磷酸氢盐、钼酸盐、碳酸盐及硝酸盐，更优选硫酸铵、磷酸铵、钼酸铵、蔗糖八硫酸酯铵、蔗糖八硫酸酯三乙铵、葡萄糖酸铜、乙基硫酸铵、乙二胺四乙酸铵盐、氯化铵、氢氧化铵、乙酸铵、葡聚糖硫酸铵或三乙基葡聚糖硫酸铵及其组合。
  8. 根据权利要求1-7任意一项所述的药物组合物，其中所述脂质体内水相中还含有至少一种pH调节剂，pH调节剂优选氢氧化钠、盐酸、氨水、磷酸、苹果酸、组氨酸、三乙醇胺。
  9. 根据权利要求1-8任意一项所述的药物组合物，其中所述式I化合物或其可药用盐的浓度选自0.01～100mg/mL，优选0.05～10mg/mL。
  10. 根据权利要求1-9任意一项所述的药物组合物，其中所述脂质体外相中含有选自糖、电解质、氨基酸或其盐中的一种或多种，优选氯化钠和组氨酸或其盐。
  11. 一种冻干组合物，其通过将权利要求1-10任意一项所述的药物组合物冷冻干燥或喷雾干燥获得。
  12. 一种冻干组合物，其复溶后可得到权利要求1-10任意一项所述的药物组合物。
  13. 一种复溶溶液，其通过将权利要求11所述的冻干组合物复溶获得。
  14. 一种包含式I化合物或其可药用盐的脂质体，
  15. 一种制备权利要求2-10任意一项所述的药物组合物或权利要求14所述脂质体的方法，所述方法包括：制备空白脂质体，以及将式I化合物或其可药用盐导入空白脂质体内水相的步骤。
  16. 权利要求1-10任意一项所述的药物组合物、权利要求11或12所述的冻干组合物或权利要求13所述的复溶溶液或权利要求14所述脂质体在制备治疗疾病或病症的药物中的用途，所述的疾病或病症优选为癌症，更优选自脑癌、甲状腺癌、头颈癌、鼻咽癌、咽喉癌、口腔癌、唾液腺癌、食道癌、胃癌、肺癌、肝癌、肾癌、胰腺癌、胆囊癌、胆管癌、结直肠癌、小肠癌、胃肠道间质瘤、尿路上皮癌、尿道癌、膀胱癌、乳腺癌、阴道癌、卵巢癌、子宫内膜癌、宫颈癌、输卵管癌、睾丸癌、前列腺癌、血管瘤、白血病、淋巴瘤、骨髓瘤、皮肤癌、脂肪瘤、骨癌、软组织肉瘤、神经纤维瘤、神经胶质瘤、成神经细胞瘤和胶质母细胞瘤。
PCT/CN2023/109901 2022-07-29 2023-07-28 一种包含kras g12d抑制剂的药物组合物 Ceased WO2024022507A1 (zh)

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Cited By (19)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US12059425B2 (en) 2022-08-05 2024-08-13 Kumquat Biosciences Inc. Heterocyclic compounds and uses thereof
WO2024206858A1 (en) 2023-03-30 2024-10-03 Revolution Medicines, Inc. Compositions for inducing ras gtp hydrolysis and uses thereof
WO2024229406A1 (en) 2023-05-04 2024-11-07 Revolution Medicines, Inc. Combination therapy for a ras related disease or disorder
WO2025034702A1 (en) 2023-08-07 2025-02-13 Revolution Medicines, Inc. Rmc-6291 for use in the treatment of ras protein-related disease or disorder
WO2025080946A2 (en) 2023-10-12 2025-04-17 Revolution Medicines, Inc. Ras inhibitors
US12338256B2 (en) 2022-05-19 2025-06-24 Genentech, Inc. Aza-tetracyclic oxazepine compounds and uses thereof
WO2025171296A1 (en) 2024-02-09 2025-08-14 Revolution Medicines, Inc. Ras inhibitors
EP4397664A4 (en) * 2021-08-31 2025-09-03 Genfleet Therapeutics Shanghai Inc CYCLIC COMPOUND FUSED TO A PYRIMIDINE, PROCESS FOR ITS PREPARATION AND ITS USE
US12448400B2 (en) 2023-09-08 2025-10-21 Gilead Sciences, Inc. KRAS G12D modulating compounds
WO2025240847A1 (en) 2024-05-17 2025-11-20 Revolution Medicines, Inc. Ras inhibitors
WO2025255438A1 (en) 2024-06-07 2025-12-11 Revolution Medicines, Inc. Methods of treating a ras protein-related disease or disorder
WO2025265060A1 (en) 2024-06-21 2025-12-26 Revolution Medicines, Inc. Therapeutic compositions and methods for managing treatment-related effects
WO2026006747A1 (en) 2024-06-28 2026-01-02 Revolution Medicines, Inc. Ras inhibitors
WO2026015825A1 (en) 2024-07-12 2026-01-15 Revolution Medicines, Inc. Use of ras inhibitor for treating pancreatic cancer
WO2026015790A1 (en) 2024-07-12 2026-01-15 Revolution Medicines, Inc. Methods of treating a ras related disease or disorder
WO2026015801A1 (en) 2024-07-12 2026-01-15 Revolution Medicines, Inc. Methods of treating a ras related disease or disorder
WO2026015796A1 (en) 2024-07-12 2026-01-15 Revolution Medicines, Inc. Methods of treating a ras related disease or disorder
WO2026050446A1 (en) 2024-08-29 2026-03-05 Revolution Medicines, Inc. Ras inhibitors
WO2026072904A2 (en) 2024-09-26 2026-04-02 Revolution Medicines, Inc. Compositions and methods for treating lung cancer

Citations (10)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN110603258A (zh) * 2017-05-11 2019-12-20 阿斯利康(瑞典)有限公司 抑制g12c突变型ras蛋白的杂芳基化合物
WO2021041671A1 (en) 2019-08-29 2021-03-04 Mirati Therapeutics, Inc. Kras g12d inhibitors
WO2021107160A1 (en) * 2019-11-29 2021-06-03 Taiho Pharmaceutical Co., Ltd. A compound having inhibitory activity against kras g12d mutation
WO2021203768A1 (zh) * 2020-04-08 2021-10-14 江苏恒瑞医药股份有限公司 嘧啶并二环类衍生物、其制备方法及其在医药上的应用
WO2022017339A1 (zh) * 2020-07-20 2022-01-27 江苏恒瑞医药股份有限公司 稠合哒嗪类衍生物、其制备方法及其在医药上的应用
WO2022188729A1 (en) * 2021-03-07 2022-09-15 Jacobio Pharmaceuticals Co., Ltd. Fused ring derivatives useful as kras g12d inhibitors
WO2022194245A1 (zh) * 2021-03-17 2022-09-22 劲方医药科技(上海)有限公司 嘧啶并环类化合物及其制法和用途
WO2022268051A1 (zh) 2021-06-21 2022-12-29 江苏恒瑞医药股份有限公司 稠合四环类化合物、其制备方法及其在医药上的应用
WO2023001123A1 (zh) * 2021-07-19 2023-01-26 上海艾力斯医药科技股份有限公司 新型吡啶并嘧啶衍生物
WO2023001141A1 (en) * 2021-07-23 2023-01-26 Shanghai Zion Pharma Co. Limited Kras g12d inhibitors and uses thereof

Family Cites Families (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US9968570B2 (en) * 2013-01-14 2018-05-15 Chemo-Enhanced Llc Compositions and methods for treating cancer
CN112745335B (zh) * 2019-10-30 2024-08-30 武汉誉祥医药科技有限公司 一种三并杂环化合物及其用途

Patent Citations (10)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN110603258A (zh) * 2017-05-11 2019-12-20 阿斯利康(瑞典)有限公司 抑制g12c突变型ras蛋白的杂芳基化合物
WO2021041671A1 (en) 2019-08-29 2021-03-04 Mirati Therapeutics, Inc. Kras g12d inhibitors
WO2021107160A1 (en) * 2019-11-29 2021-06-03 Taiho Pharmaceutical Co., Ltd. A compound having inhibitory activity against kras g12d mutation
WO2021203768A1 (zh) * 2020-04-08 2021-10-14 江苏恒瑞医药股份有限公司 嘧啶并二环类衍生物、其制备方法及其在医药上的应用
WO2022017339A1 (zh) * 2020-07-20 2022-01-27 江苏恒瑞医药股份有限公司 稠合哒嗪类衍生物、其制备方法及其在医药上的应用
WO2022188729A1 (en) * 2021-03-07 2022-09-15 Jacobio Pharmaceuticals Co., Ltd. Fused ring derivatives useful as kras g12d inhibitors
WO2022194245A1 (zh) * 2021-03-17 2022-09-22 劲方医药科技(上海)有限公司 嘧啶并环类化合物及其制法和用途
WO2022268051A1 (zh) 2021-06-21 2022-12-29 江苏恒瑞医药股份有限公司 稠合四环类化合物、其制备方法及其在医药上的应用
WO2023001123A1 (zh) * 2021-07-19 2023-01-26 上海艾力斯医药科技股份有限公司 新型吡啶并嘧啶衍生物
WO2023001141A1 (en) * 2021-07-23 2023-01-26 Shanghai Zion Pharma Co. Limited Kras g12d inhibitors and uses thereof

Non-Patent Citations (2)

* Cited by examiner, † Cited by third party
Title
KARGBO ROBERT B.: "Targeting KRAS G12D Mutations: Discovery of Small Molecule Inhibitors for the Potential Treatment of Intractable Cancers", ACS MEDICINAL CHEMISTRY LETTERS, AMERICAN CHEMICAL SOCIETY, US, vol. 14, no. 8, 10 August 2023 (2023-08-10), US , pages 1041 - 1042, XP093133604, ISSN: 1948-5875, DOI: 10.1021/acsmedchemlett.3c00277 *
See also references of EP4563151A4

Cited By (19)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
EP4397664A4 (en) * 2021-08-31 2025-09-03 Genfleet Therapeutics Shanghai Inc CYCLIC COMPOUND FUSED TO A PYRIMIDINE, PROCESS FOR ITS PREPARATION AND ITS USE
US12338256B2 (en) 2022-05-19 2025-06-24 Genentech, Inc. Aza-tetracyclic oxazepine compounds and uses thereof
US12059425B2 (en) 2022-08-05 2024-08-13 Kumquat Biosciences Inc. Heterocyclic compounds and uses thereof
WO2024206858A1 (en) 2023-03-30 2024-10-03 Revolution Medicines, Inc. Compositions for inducing ras gtp hydrolysis and uses thereof
WO2024229406A1 (en) 2023-05-04 2024-11-07 Revolution Medicines, Inc. Combination therapy for a ras related disease or disorder
WO2025034702A1 (en) 2023-08-07 2025-02-13 Revolution Medicines, Inc. Rmc-6291 for use in the treatment of ras protein-related disease or disorder
US12448400B2 (en) 2023-09-08 2025-10-21 Gilead Sciences, Inc. KRAS G12D modulating compounds
WO2025080946A2 (en) 2023-10-12 2025-04-17 Revolution Medicines, Inc. Ras inhibitors
WO2025171296A1 (en) 2024-02-09 2025-08-14 Revolution Medicines, Inc. Ras inhibitors
WO2025240847A1 (en) 2024-05-17 2025-11-20 Revolution Medicines, Inc. Ras inhibitors
WO2025255438A1 (en) 2024-06-07 2025-12-11 Revolution Medicines, Inc. Methods of treating a ras protein-related disease or disorder
WO2025265060A1 (en) 2024-06-21 2025-12-26 Revolution Medicines, Inc. Therapeutic compositions and methods for managing treatment-related effects
WO2026006747A1 (en) 2024-06-28 2026-01-02 Revolution Medicines, Inc. Ras inhibitors
WO2026015825A1 (en) 2024-07-12 2026-01-15 Revolution Medicines, Inc. Use of ras inhibitor for treating pancreatic cancer
WO2026015790A1 (en) 2024-07-12 2026-01-15 Revolution Medicines, Inc. Methods of treating a ras related disease or disorder
WO2026015801A1 (en) 2024-07-12 2026-01-15 Revolution Medicines, Inc. Methods of treating a ras related disease or disorder
WO2026015796A1 (en) 2024-07-12 2026-01-15 Revolution Medicines, Inc. Methods of treating a ras related disease or disorder
WO2026050446A1 (en) 2024-08-29 2026-03-05 Revolution Medicines, Inc. Ras inhibitors
WO2026072904A2 (en) 2024-09-26 2026-04-02 Revolution Medicines, Inc. Compositions and methods for treating lung cancer

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JP2025524799A (ja) 2025-08-01
TW202408536A (zh) 2024-03-01
EP4563151A1 (en) 2025-06-04
CN119384279A (zh) 2025-01-28
EP4563151A4 (en) 2025-11-05

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