WO2023123986A1 - 优替德隆脂质体组合物及其制备方法和用途 - Google Patents

优替德隆脂质体组合物及其制备方法和用途 Download PDF

Info

Publication number
WO2023123986A1
WO2023123986A1 PCT/CN2022/103468 CN2022103468W WO2023123986A1 WO 2023123986 A1 WO2023123986 A1 WO 2023123986A1 CN 2022103468 W CN2022103468 W CN 2022103468W WO 2023123986 A1 WO2023123986 A1 WO 2023123986A1
Authority
WO
WIPO (PCT)
Prior art keywords
liposome composition
phospholipids
liposome
phospholipid
composition according
Prior art date
Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
Ceased
Application number
PCT/CN2022/103468
Other languages
English (en)
French (fr)
Inventor
唐莉
张川
徐靖
张�成
邱荣国
Current Assignee (The listed assignees may be inaccurate. Google has not performed a legal analysis and makes no representation or warranty as to the accuracy of the list.)
Chengdu Biostar Pharmaceuticals Ltd
Original Assignee
Chengdu Biostar Pharmaceuticals Ltd
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Application filed by Chengdu Biostar Pharmaceuticals Ltd filed Critical Chengdu Biostar Pharmaceuticals Ltd
Priority to US18/264,506 priority Critical patent/US20240108579A1/en
Priority to CN202280010753.3A priority patent/CN116710085A/zh
Priority to EP22913275.8A priority patent/EP4275676B1/en
Priority to JP2023542005A priority patent/JP2024505154A/ja
Publication of WO2023123986A1 publication Critical patent/WO2023123986A1/zh
Anticipated expiration legal-status Critical
Ceased legal-status Critical Current

Links

Images

Classifications

    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K9/00—Medicinal preparations characterised by special physical form
    • A61K9/10—Dispersions; Emulsions
    • A61K9/127—Synthetic bilayered vehicles, e.g. liposomes or liposomes with cholesterol as the only non-phosphatidyl surfactant
    • A61K9/1271—Non-conventional liposomes, e.g. PEGylated liposomes or liposomes coated or grafted with polymers
    • 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
    • A61K9/1274—Non-vesicle bilayer structures, e.g. liquid crystals, tubules, cubic phases or cochleates; Sponge phases
    • 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
    • A61K31/41—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having five-membered rings with two or more ring hetero atoms, at least one of which being nitrogen, e.g. tetrazole
    • A61K31/425—Thiazoles
    • A61K31/426—1,3-Thiazoles
    • 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
    • A61K31/41—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having five-membered rings with two or more ring hetero atoms, at least one of which being nitrogen, e.g. tetrazole
    • A61K31/425—Thiazoles
    • A61K31/427—Thiazoles not condensed and containing further heterocyclic rings
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K45/00—Medicinal preparations containing active ingredients not provided for in groups A61K31/00 - A61K41/00
    • A61K45/06—Mixtures of active ingredients without chemical characterisation, e.g. antiphlogistics and cardiaca
    • 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
    • A61K9/10—Dispersions; Emulsions
    • A61K9/127—Synthetic bilayered vehicles, e.g. liposomes or liposomes with cholesterol as the only non-phosphatidyl surfactant
    • A61K9/1271—Non-conventional liposomes, e.g. PEGylated liposomes or liposomes coated or grafted with polymers
    • A61K9/1272—Non-conventional liposomes, e.g. PEGylated liposomes or liposomes coated or grafted with polymers comprising non-phosphatidyl surfactants as bilayer-forming substances, e.g. cationic lipids or non-phosphatidyl liposomes coated or grafted with polymers
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P35/00—Antineoplastic agents

Definitions

  • the invention relates to the technical field of medicine, and more specifically relates to a liposome composition containing eutendrone (or its salt), phospholipid and optional sterol, and a preparation method and application thereof.
  • Utilidelon is a class of epothilone derivatives, a macrolide compound, and a secondary metabolite produced by the genetically modified myxobacteria S. cellulosus. Studies have shown that epothilone antibiotics have the same pharmacological mechanism as paclitaxel, which exerts anti-tumor effects by inhibiting the depolymerization of tubulin.
  • the chemical name of Youtedron is: 4,8-dihydroxy-5,5,7,9,13-pentamethyl-16-[1-methyl-2-(2-methyl-thiazole-4- base)-vinyl]-hexadecane-13-en-2,6-one lactone.
  • Utenidron is easily soluble in ethanol, methanol, ethyl acetate, and chloroform, but hardly soluble in water.
  • the saturated solubility in water is lower than 1 ⁇ g/ml.
  • the clinical formulations contain a large amount of polyoxyethylene (35) Castor oil and organic solvents have the possibility of causing severe allergic reactions. Before clinical application, patients need to be given preventive anti-allergic treatment, and patients' medication compliance is poor.
  • one of the technical problems to be solved by the present invention is to provide a stable euteron liposome.
  • the technical problem to be solved by the present invention is to provide a stable eutedrone liposome with high drug loading.
  • the technical problem to be solved by the present invention is to provide a method for preparing stable utidron liposomes, which can stably encapsulate utidron in liposomes with high efficiency.
  • the inventors creatively used the special physical and chemical properties of different types of phospholipids, selected phospholipids or phospholipid combinations suitable for eutenedron, and prepared one or more stable eutedron liposomes.
  • the present invention provides a liposome composition, wherein the liposome composition comprises eutendron and phospholipids.
  • the present invention also provides a liposome composition, wherein the liposome composition comprises eutendrone, phospholipid and sterol.
  • Ustedron is entrapped in the liposome.
  • the phospholipid is selected from one or more of pegylated phospholipids, anionic phospholipids, cationic phospholipids and zwitterionic phospholipids.
  • the phospholipid comprises or consists of a zwitterionic phospholipid.
  • the phospholipids comprise or consist of zwitterionic phospholipids in combination with pegylated phospholipids, anionic phospholipids and/or cationic phospholipids.
  • the phospholipid comprises or consists of a combination of zwitterionic phospholipids and pegylated phospholipids. In one or more embodiments, the phospholipid comprises or consists of a combination of zwitterionic and anionic phospholipids. In one or more embodiments, the phospholipid comprises or consists of a combination of a zwitterionic phospholipid and a cationic phospholipid. In one or more embodiments, the phospholipids comprise or consist of zwitterionic phospholipids in combination with pegylated phospholipids and anionic phospholipids. In one or more embodiments, the phospholipid comprises or consists of a zwitterionic phospholipid in combination with a pegylated phospholipid and a cationic phospholipid.
  • the zwitterionic phospholipids are selected from egg phosphatidylcholine (EPC), egg phosphatidylserine (EPS), phosphatidylethanolamine (EPE), soybean phosphatidylserine (SPS), soybean phospholipids Ethanolamine (SPE), hydrogenated egg phosphatidylcholine (HEPC), hydrogenated egg phosphatidylserine (HEPS), hydrogenated egg phosphatidylethanolamine (HEPE), hydrogenated soybean phosphatidylcholine (HSPC), hydrogenated soybean phosphatidylserine ( HSPS), hydrogenated soybean phosphatidylethanolamine (HSPE), dipalmitoylphosphatidylcholine (DPPC), 1-palmitoyl-2-myristoylphosphatidylcholine (PMPC), 1-myristoyl-2-palmitoylcholine Acylphosphatidylcholine (MPPC), Dioleoylphosphati
  • the zwitterionic phospholipids are selected from unsaturated zwitterionic phospholipids.
  • unsaturated zwitterionic phospholipids include, but are not limited to, egg phosphatidylcholine (EPC), egg phosphatidylserine (EPS), phosphatidylethanolamine (EPE), soybean phosphatidylserine (SPS), soybean phosphatidylethanolamine (SPE) , dioleoylphosphatidylcholine (DOPC), 1,2-docosenoyl-sn-glyceryl-3-phosphocholine (DEPC), palmitoyloleoylphosphatidylcholine (POPC), two Oleoylphosphatidylethanolamine (DOPE), palmitoyloleoylphosphatidylethanolamine (POPE).
  • EPC egg phosphatidylcholine
  • EPS egg phosphatidylserine
  • EPE phosphatidyl
  • the zwitterionic phospholipids are selected from synthetic zwitterionic phospholipids.
  • synthetic zwitterionic phospholipids include, but are not limited to, DSPC and DOPC.
  • the zwitterionic phospholipids are selected from unsaturated synthetic zwitterionic phospholipids.
  • unsaturated synthetic zwitterionic phospholipids include, but are not limited to, DOPC.
  • the zwitterionic phospholipids are selected from the group consisting of egg phosphatidylcholine (EPC), hydrogenated soybean phosphatidylcholine (HSPC), dioleoylphosphatidylcholine (DOPC), distearyl One or more of acylphosphatidylcholines (DSPC).
  • EPC egg phosphatidylcholine
  • HSPC hydrogenated soybean phosphatidylcholine
  • DOPC dioleoylphosphatidylcholine
  • DSPC distearyl
  • acylphosphatidylcholines acylphosphatidylcholines
  • the zwitterionic phospholipids are selected from the group consisting of egg phosphatidylcholine (EPC), dioleoylphosphatidylcholine (DOPC), and distearoylphosphatidylcholine (DSPC). one or more.
  • EPC egg phosphatidylcholine
  • DOPC dioleoylphosphatidylcholine
  • DSPC distearoylphosphatidylcholine
  • the zwitterionic phospholipid is selected from one or both of egg phosphatidylcholine (EPC) and dioleoylphosphatidylcholine (DOPC).
  • EPC egg phosphatidylcholine
  • DOPC dioleoylphosphatidylcholine
  • the zwitterionic phospholipid is dioleoylphosphatidylcholine (DOPC).
  • DOPC dioleoylphosphatidylcholine
  • the pegylated phospholipid is selected from distearoylphosphatidylethanolamine-polyethylene glycol (DSPE-PEG), distearoylphosphatidylethanolamine-methoxypolyethylene glycol alcohol (DSPE-MPEG), dimyristoylphosphatidylethanolamine-polyethylene glycol (DMPE-PEG), dipalmitoylglycerylsuccinate polyethylene glycol (DPGS-PEG), cholesteryl-pegylated phospholipids and one or more of ceramide-based pegylated phospholipids.
  • DSPE-PEG distearoylphosphatidylethanolamine-polyethylene glycol
  • DSPE-MPEG distearoylphosphatidylethanolamine-methoxypolyethylene glycol alcohol
  • DMPE-PEG dimyristoylphosphatidylethanolamine-polyethylene glycol
  • DPGS-PEG dipalmitoylglycerylsuccinate polyethylene
  • the pegylated phospholipids are selected from distearoylphosphatidylethanolamine-polyethylene glycol (DSPE-PEG) and distearoylphosphatidylethanolamine-methoxypolyethylene glycol One or both of diols (DSPE-MPEG).
  • the pegylated phospholipid is distearoylphosphatidylethanolamine-methoxypolyethylene glycol (DSPE-MPEG).
  • the anionic phospholipid is selected from di(hexadecyl) phosphate (DhP), phosphatidylinositol, phosphatidylserine, phosphatidylglycerol, lysylphosphatidylglycerol (LPG) , phosphatidylethanolamine, phosphatidic acid, cardiolipin and cholesterol hemisuccinate one or more.
  • DhP di(hexadecyl) phosphate
  • phosphatidylinositol phosphatidylserine
  • phosphatidylglycerol lysylphosphatidylglycerol (LPG)
  • phosphatidylethanolamine phosphatidic acid
  • cardiolipin cardiolipin
  • cholesterol hemisuccinate one or more.
  • the phosphatidylserine is selected from one or more of dimyristoylphosphatidylserine, dipalmitoylphosphatidylserine and distearoylphosphatidylserine.
  • the anionic phospholipids are selected from phosphatidylglycerols.
  • the phosphatidylglycerol is selected from dimyristoylphosphatidylglycerol (DMPG), dipalmitoylphosphatidylglycerol (DPPG), distearoylphosphatidylglycerol (DSPG), dioleic acid Acylphosphatidylglycerol (DOPG), Dilauroylphosphatidylglycerol (DLPG), Egg Phosphatidylglycerol (EPG), Egg Phosphatidylinositol (EPI), Soybean Phosphatidylglycerol (SPG), Soybean Phosphatidylinositol (SPI) ), hydrogenated lecithin phosphatidylglycerol (HEPG), hydrogenated soybean phosphatidyl glycerol (HSPG), hydrogenated lecithin phosphatidylinositol (HEPI), and palmi
  • DMPG dim
  • the phosphatidylglycerol is selected from distearoylphosphatidylglycerol (DSPG).
  • the cationic phospholipid is selected from N-[1-(2,3-dioleoyloxy)propyl]-N,N,N-trimethylammonium salt (DOTAP), N-[1-(2,3-Dimyristoyl)propyl]-N,N,N-trimethylammonium salt (DMTAP), N-[1-(2,3-Dipalmitoyl)propyl ]-N,N,N-trimethylammonium salt (DPTAP), N-[1-(2,3-distearoyl)propyl]-N,N,N-trimethylammonium salt (DSTAP) , Dimethyl di(octadecyl)ammonium bromide (DDAB), 1,2-diacyloxy-3-trimethylammonium propane, N-[1-(2,3-dioleoyloxy )Propyl]-N,N-Dimethylamine (DODAP), 1,2-Di
  • the phospholipid comprises or consists of a combination of unsaturated zwitterionic phospholipids and anionic phospholipids. In one or more embodiments, the phospholipid comprises or consists of a combination of synthetic zwitterionic phospholipids and anionic phospholipids. In one or more preferred embodiments, the phospholipids comprise or consist of unsaturated synthetic zwitterionic phospholipids in combination with anionic phospholipids. In one or more more preferred embodiments, the phospholipid comprises or consists of a combination of dioleoylphosphatidylcholine (DOPC) and distearoylphosphatidylglycerol (DSPG).
  • DOPC dioleoylphosphatidylcholine
  • DSPG distearoylphosphatidylglycerol
  • the phospholipid comprises or consists of a combination of unsaturated zwitterionic phospholipids and pegylated phospholipids. In one or more embodiments, the phospholipid comprises or consists of a combination of synthetic zwitterionic phospholipids and pegylated phospholipids. In one or more preferred embodiments, the phospholipid comprises or consists of a combination of unsaturated synthetic zwitterionic phospholipids and pegylated phospholipids.
  • the phospholipid comprises a combination of dioleoylphosphatidylcholine (DOPC) and distearoylphosphatidylethanolamine-methoxypolyethylene glycol (DSPE-MPEG) or is derived from The combination consists of.
  • DOPC dioleoylphosphatidylcholine
  • DSPE-MPEG distearoylphosphatidylethanolamine-methoxypolyethylene glycol
  • the sterol is selected from one of cholesterol, 7-hydrocholesterol, lanosterol, sitosterol, brassicasterol, mycosterol, oysterol, stigmasterol, and ergosterol or more. In one or more preferred embodiments, the sterol is selected from one or both of cholesterol and ergosterol. In one or more more preferred embodiments, the sterol is cholesterol.
  • the liposome composition comprises eutendrone and zwitterionic phospholipids, such as zwitterionic phospholipids as described above, preferably selected from one or more of EPC, DOPC and DSPC . In one or more more preferred embodiments, the liposome composition comprises eutendrone and DOPC.
  • the liposome composition comprises eutendrone, a zwitterionic phospholipid, and a pegylated phospholipid.
  • the liposome composition comprises Utenidron, DSPE-MPEG and zwitterionic phospholipids, wherein the zwitterionic phospholipids are zwitterionic phospholipids as described above, preferably selected from HSPC, One or more of EPC, DOPC and DSPC, preferably selected from one or more of HSPC, EPC and DOPC.
  • the liposome composition comprises eutendrone, zwitterionic phospholipids and sterols. In one or more more preferred embodiments, the liposome composition comprises euteron, zwitterionic phospholipids and cholesterol, wherein the zwitterionic phospholipids are zwitterionic phospholipids as described above, preferably selected from HSPC, One or more of EPC, DOPC and DSPC, more preferably one or more selected from EPC, DOPC and DSPC. In one or more more preferred embodiments, the liposome composition comprises eutendrone, EPC and cholesterol. In one or more more preferred embodiments, the liposome composition comprises eutendrone, DOPC, DSPC and cholesterol.
  • the liposome composition comprises eutendrone, pegylated phospholipids, zwitterionic phospholipids and sterols.
  • the liposome composition comprises Utenidron, DSPE-MPEG, zwitterionic phospholipids and cholesterol, wherein the zwitterionic phospholipids are zwitterionic phospholipids as described above, preferably One or more of DOPC, DSPC, HSPC and EPC, more preferably one or more of DOPC, HSPC and EPC.
  • the liposome composition comprises eutendrone, DSPE-MPEG, DOPC and cholesterol.
  • the liposome composition comprises eutendrone, anionic phospholipids, zwitterionic phospholipids and sterols. In one or more preferred embodiments, the liposome composition comprises euteron, DSPG, zwitterionic phospholipids and cholesterol, wherein the zwitterionic phospholipids are zwitterionic phospholipids as described above, preferably selected from DOPC One or more of , DSPC, HSPC and EPC, more preferably selected from one or more of DOPC, DSPC and EPC. In one or more more preferred embodiments, the liposome composition comprises eutendrone, DSPG, DOPC and cholesterol.
  • the liposome composition does not include a liposome composition consisting only of eutilone and HSPC.
  • the content of eutendron is 2.5-20% by weight, preferably 3-10% by weight .
  • the content of the phospholipid is 40.0-97.5% by weight, preferably 65.0-97.0% by weight.
  • the content of the sterol when sterol is contained, based on the total weight of the liposome composition, is 0.8-48.8% by weight, preferably 0.9% by weight. - 48.5% by weight, more preferably 3-30% by weight.
  • the phospholipid when the phospholipid is a combination of zwitterionic phospholipids and pegylated phospholipids, anionic phospholipids and/or cationic phospholipids, according to the total weight of phospholipids Calculated, the content of the zwitterionic phospholipid is 50-90% by weight; the content of the pegylated phospholipid is 0-40% by weight; the content of the anionic phospholipid is 0-40% by weight; the content of the cationic phospholipid The content is 0-40% by weight.
  • the phospholipid when the phospholipid is a combination of zwitterionic phospholipids and pegylated phospholipids, based on the total weight of phospholipids, the zwitterionic phospholipids
  • the content is 60%-90%, preferably 70%-85%; the content of the pegylated phospholipid is 10-40%, preferably 15%-30%.
  • the phospholipid when the phospholipid is a combination of zwitterionic phospholipids and anionic phospholipids, based on the total weight of phospholipids, the content of the zwitterionic phospholipids is 60 %-90%, preferably 65%-85%, more preferably 70%-80%; the content of the anionic phospholipid is 10-40%, preferably 15%-35%, more preferably 20%-30% .
  • the phospholipid when the phospholipid is a combination of zwitterionic phospholipids and cationic phospholipids, the content of the zwitterionic phospholipids is 60% by total weight of phospholipids %-90%, preferably 65%-85%, more preferably 70%-80%; the content of the cationic phospholipid is 10-40%, preferably 15%-35%, more preferably 20%-30% .
  • the mass ratio of eutilone to total phospholipids is 0.2% to 30%, preferably 2.6-25.3%, more preferably 2.6%-11.2%, more preferably 3.0-8.0%.
  • the mass ratio of sterols to total phospholipids is 1%-50%, preferably 3%-40%, more preferably 5%-35% .
  • the liposome composition further comprises one or more of an osmotic pressure regulator, an antioxidant, a preservative, a pH regulator, and a buffer.
  • the osmotic pressure regulator is selected from one or more of sodium chloride, glycerin, sorbitol, mannitol and glucose.
  • the pH regulator is selected from one or more of sodium hydroxide, sodium citrate, citric acid, phosphoric acid, acetic acid and hydrochloric acid.
  • the preservative is selected from one or more of alkyl hydroxybenzoate, benzoic acid, sodium benzoate, sorbic acid, chlorethyl acetate and benzalkonium bromide.
  • the antioxidant is selected from sodium sulfite, sodium bisulfite, sodium metabisulfite, sodium thiosulfate, ascorbic acid, tert-butyl p-hydroxyanisole, 2,6-di-tert-butylated hydroxy One or more of toluene and vitamin E.
  • the buffer is selected from one or more of citrate buffer, phosphate buffer, acetate buffer and Tris buffer.
  • the liposome composition further comprises one of deionized water, 10% sucrose aqueous solution, phosphate buffer, and citrate buffer.
  • the liposome composition is in liquid form.
  • concentration of eutilone in the liposome composition in liquid form is 0.01 mg/ml-20 mg/ml, preferably 0.5 mg/ml-5 mg/ml.
  • the liposome composition is in the form of a lyophilized powder.
  • the liposome composition in the form of a lyophilized powder may further contain a lyoprotectant.
  • the lyoprotectant is selected from one or more of glucose, sucrose, maltose, lactose, mannose, trehalose, glycine, and dextran.
  • the particle size of liposomes in the liposome composition is less than 250 nm, preferably 50 nm to 220 nm.
  • the average polydispersity index (PDI) of the liposomes in the liposome composition is less than 0.3, preferably less than 0.2.
  • the liposome composition of the present invention can be selected from shear mixing method, film hydration method, spray drying method, freeze drying method, freeze-thaw method, solvent injection method, reverse phase evaporation Method, emulsification volatilization method, microfluidic method, ultrasonic method, supercritical fluid method, homogeneous method in one or more methods to prepare.
  • the liposome composition of the present invention can be prepared by thin film hydration method.
  • the membrane hydration method comprises the following steps:
  • step (3) hydrating the lipid film obtained in step (2) to obtain a liposome solution
  • step (3) The liposome solution obtained in step (3) is sized to obtain a nanoliposome solution.
  • the solvent used in the step (1) is an organic solvent or a mixture of an organic solvent and water.
  • the organic solvent is selected from chloroform, dichloromethane, tert-butanol, isopropanol, ethyl acetate, ethanol, methanol, tetrahydrofuran, dioxane, acetonitrile, acetone, dimethyl One or more of sulfoxide, dimethylformamide, and methylpyrrolidone.
  • the solvent used in the step (1) is selected from a mixture of dichloromethane and methanol; a mixture of dichloromethane and ethanol; and a mixture of chloroform, methanol and water.
  • the solvent used in the step (1) is selected from the mixture of dichloromethane and methanol 5:1 or 6:1; the mixture of dichloromethane and ethanol 5:1; and chloroform, Methanol and water 95:4:1 mixture.
  • a hydration solvent is used to hydrate the lipid film.
  • the hydration solvent is one or more of deionized water, 10% sucrose aqueous solution, phosphate buffer, and citrate buffer.
  • the hydration solvent further contains one or more of osmotic pressure regulators, antioxidants, preservatives, pH regulators, and buffers.
  • the sizing is carried out using one or more methods selected from the group consisting of shearing method, high-pressure homogenization method and microfluidic homogenization method.
  • the film hydration method further includes step (5): sterilizing the nanoliposome solution obtained in step (4).
  • the film hydration method further includes the step of lyophilizing the drug solution obtained in step (5).
  • the present invention provides the use of the liposome composition described in the present invention in the preparation of medicaments for preventing or treating cancer.
  • the cancer is a solid tumor, such as breast cancer, lung cancer, and digestive tract tumors.
  • the liposome composition is prepared as a pharmaceutical formulation.
  • the liposomal composition is formulated parenterally, by inhalation, intraperitoneally, intravesically, intramuscularly, intravenously, intratracheally, subcutaneously, intraocularly, intrathecally, Pharmaceutical formulations for transdermal, rectal or intravaginal administration.
  • the liposome composition is prepared as a pharmaceutical preparation for intravenous administration.
  • the pharmaceutical preparation is a solid preparation, a liquid preparation or a gas preparation, more preferably a liquid preparation for injection.
  • the pharmaceutical preparation is a stable aqueous suspension reconstituted from the sterile lyophilized powder of the liposome composition before use.
  • the pharmaceutical formulation prepared from the liposomal composition also contains other drugs.
  • the other drug is an anticancer drug.
  • the invention provides a stable uteridone liposome composition.
  • the Utenidron liposome composition of the present invention does not contain excipients that are likely to cause allergic reactions in the human body, and can effectively eliminate the allergic reactions caused by castor oil and the irritant reactions caused by organic solvents in existing clinical preparations, thereby increasing the drug use of patients compliance, while prolonging the half-life of Utenidron in vivo, to achieve the clinical effect of attenuating toxicity and increasing efficacy.
  • the formula combination adopted by the present invention can increase the weight ratio of eutilone in the liposome to more than 3%.
  • the increase of this amount means the increase of the drug loading, and the drug loading of the liposome of the present application can reach 3%. %, even as high as 7%, which is much higher than the drug loading capacity of the existing Utenidron preparations.
  • Increased drug loading can reduce the amount of excipients, reduce production costs, reduce the burden of medication for patients, and at the same time reduce the possible side effects of excipients .
  • the Utenidron liposome composition of the present invention has high drug loading capacity and stability, and has good industrialization potential.
  • Fig. 1 shows the hydration particle size distribution figure of the Utenidron liposome prepared in Example 1.
  • Fig. 2 represents the test result of the in vitro release degree of the Utilidelon liposome prepared in Example 5.
  • Fig. 3 shows the hydration particle size distribution figure of the Utenidron liposome prepared in Example 6.
  • Fig. 4 shows the hydration particle size distribution diagram of the Utenidron liposome prepared in Example 10.
  • Fig. 5 shows the morphological characterization diagram of the Utenidron liposome prepared in Example 17.
  • the content determination was carried out according to the high performance liquid chromatography (Chinese Pharmacopoeia 2020 Edition Sibu General Rules 0512).
  • Sample tray temperature 25°C
  • Reference substance solution Dilute the stock solution of Utilidelon reference substance with acetonitrile to make a series of concentrations: 10 ⁇ g/ml, 20 ⁇ g/ml, 50 ⁇ g/ml, 100 ⁇ g/ml, 200 ⁇ g/ml.
  • test solution take the sample to be tested, add appropriate amount of 75% isopropanol and shake to redissolve, transfer all the solution to a 10ml volumetric flask, then rinse the sample bottle with 75% isopropanol, and dissolve the rinse solution Transfer to a volumetric flask to constant volume, shake well, and filter with a 0.22 ⁇ m organic filter.
  • Test example 4 Determination of the encapsulation efficiency of Youteidelong liposome
  • test solution (encapsulated dose): take 0.2ml of the Utenidron liposome solution to be tested and put it on the G-50 column of dextran fat coagulation column for separation, first use 30ml of PBS (6.8) eluent Carry out elution, collect the first 15ml of eluate, measure the content of eutendron in it as the encapsulated drug amount. Then use 30ml of physiological sodium chloride solution to elute the remaining free drug.
  • Need testing solution (medicine total amount) preparation get the eutenedron liposome solution 0.2ml to be tested in addition, dissolve and dilute to 10ml with 60% acetonitrile, shake up, filter, measure wherein the content of eutedron is the total amount of encapsulated and unencapsulated drug.
  • the encapsulation efficiency is calculated according to the following formula:
  • Encapsulation efficiency (%) amount of drug encapsulated in liposome/total amount of drug ⁇ 100.
  • the release testing method is as follows:
  • Control solution Weigh an appropriate amount of Utendelon reference substance, weigh it accurately, put it in a volumetric flask, dissolve it with a blank solution and dilute it to 0.05 ⁇ g/ml.
  • Preparation of the test solution Precisely draw 0.5ml of the Utenidron liposome solution to be tested, place it in a dialysis bag, and after two sections are tied tightly, use 500ml of phosphate buffered saline (pH7.4) as the release medium, and the rotation speed 100 rpm, temperature 37 ⁇ 0.5°C, take and release at 0.5h, 1.0h, 2.0h, 3.0h, 4.0h, 6.0h, 10.0h, 14.0h, 18.0h, 20.0h, 22.0h, 24.0h medium.
  • phosphate buffered saline pH7.4
  • the initial filtrate was 4ml, the sampling volume was 8ml, and 12.0mL of blank release medium was supplemented at the same time, 1ml of the sample at each sampling point was accurately taken in a beaker, and then 1ml of acetonitrile was added to mix, filtered, processed and injected for analysis. After the system is equilibrated, samples are injected sequentially, the chromatograms are recorded and the test results are recorded in the relevant records. Calculate the cumulative release according to the following formula.
  • a s the peak area of Utilidelone in the test solution
  • V the dilution factor of the test solution, 1000
  • a r the average peak area of the control solution Utidelon
  • M the total amount of Utilideron input in the release medium
  • V 1 sample volume for release test, 12mL
  • V 2 volume of dissolution medium, 500mL.
  • Mobile phase A methanol-tetrahydrofuran-0.17mol/L ammonium acetate aqueous solution (89:10:1)
  • mobile phase B 4mM ammonium acetate aqueous solution
  • chromatographic column Narochrom chromcore 120 C18 (3 ⁇ m, 4.6 ⁇ 150mm)
  • column temperature 30°C drift tube temperature 45°C, atomization temperature 36°C, gas flow rate 40psi, gain 100.
  • Reference substance solution Accurately weigh phospholipids or sterols, set appropriate amount, dissolve and dilute with methanol to form a series of linear control solutions.
  • Need testing solution take 0.5ml of Utilidelon liposome solution to be tested and put it in a 10ml volumetric flask, dissolve it by ultrasonic demulsification with methanol, dilute to the mark, filter through a microporous membrane of 0.45 ⁇ m, and set aside.
  • Utenidron liposome drug loading Utenidron content/(Utenidron content + total phospholipid content + sterol content)
  • the liposome solution is filtered using a 0.22 ⁇ m polyethersulfone filter membrane, and the dynamic light scattering (DLS ) to detect liposome hydration particle size.
  • the particle size diagram is shown in Figure 1, the average particle size is 181.3nm, and the PDI is 0.067.
  • the Utenidron liposomes prepared by the present invention have extremely high stability, and the drug content is still close to 100% when stored at 2-8°C for up to two months, and the particle size and particle size distribution did not change significantly.
  • the hydrated particle size of the plastid is 61.33nm
  • the polydispersity index (PDI) is 0.168
  • the content of Utilidelon is 0.127mg/ml.
  • the encapsulation rate is greater than 90%.
  • the liposome solution is filtered with a 0.22 ⁇ m polyethersulfone filter membrane, and the liposome hydration particle size is 144.5 nm detected by dynamic light scattering (DLS).
  • the dispersion index (PDI) was 0.069.
  • the particle size diagram is shown in Figure 3. The encapsulation efficiency was 94.3%.
  • the hydration solvent (10% sucrose solution, 3ml) at 50-55°C hydrates the lipid film, and after hydration, use an extruder (Avanti) to extrude the hydration solution, the extruded film is 100nm, and the number of extrusions For 10-20 times, use a 0.22 ⁇ m polyethersulfone filter membrane to filter the liposome solution after extrusion, use dynamic light scattering (DLS) to detect liposome hydration particle size is 193.2nm, polydispersity index (PDI ) was 0.143, and the content of Utenidron was 1.624mg/ml. The encapsulation rate is greater than 90%.
  • the liposome solution is filtered with a 0.22 ⁇ m polyethersulfone filter membrane, and the liposome hydration particle size is detected by dynamic light scattering (DLS) It is 196.9nm, the polydispersity index (PDI) is 0.066, and the content of Utilidelon is 1.717mg/ml.
  • DLS dynamic light scattering
  • PDI polydispersity index
  • the encapsulation rate is greater than 90%.
  • the liposome solution is filtered using a 0.22 ⁇ m polyethersulfone filter membrane, and the liposome hydration particle size is detected by dynamic light scattering (DLS). 155.8nm, polydispersity index (PDI) is 0.079. The encapsulation rate is greater than 90%.
  • DLS dynamic light scattering
  • PDI polydispersity index
  • the liposome solution is filtered using a 0.22 ⁇ m polyethersulfone filter membrane, and the liposome hydration is detected by dynamic light scattering (DLS).
  • the particle size is 136.9nm, and the polydispersity index (PDI) is 0.101.
  • the particle size diagram is shown in Figure 4. The encapsulation efficiency is greater than 90%.
  • the liposome solution is filtered using a 0.22 ⁇ m polyethersulfone filter membrane, and the liposome hydration is detected by dynamic light scattering (DLS).
  • the particle size is 201.9nm
  • the polydispersity index (PDI) is 0.188
  • the content of eutilidelon is 2.061mg/ml
  • the encapsulation efficiency is greater than 90%.
  • the liposome solution is filtered with a 0.22 ⁇ m polyethersulfone filter membrane, and the liposome solution is detected using dynamic light scattering (DLS).
  • the hydrated particle size of the plastid is 147.3nm, and the polydispersity index (PDI) is 0.155.
  • the encapsulation rate is greater than 90%.
  • the liposome solution is filtered with a 0.22 ⁇ m polyethersulfone filter membrane, and the lipid is detected by dynamic light scattering (DLS).
  • the body hydration particle size is 134.3nm, and the polydispersity index (PDI) is 0.11.
  • the encapsulation rate is greater than 90%.
  • the liposome solution is filtered with a 0.22 ⁇ m polyethersulfone filter membrane, and the filtered medicinal solution is divided into cillin Put it into the bottle and put it into a lyophilizer for lyophilization.
  • the lyophilization parameters are: pre-freeze at -45°C for 120 minutes, heat up to -30°C and then dry for 1000 minutes, and heat up to 25°C for 720 minutes.
  • DLS dynamic light scattering
  • PDI polydispersity index
  • the encapsulation rate is greater than 90%.
  • freeze-drying is carried out in a dryer, and the freeze-drying parameters are: pre-freeze at -45°C for 120 minutes, heat up to -30°C and then dry for 1,000 minutes for the first time, and heat up to 25°C for 720 minutes for the second time.
  • DLS dynamic light scattering
  • PDI polydispersity index
  • eutilon was 0.799 mg/ml. The encapsulation rate is greater than 90%.
  • the extruded film is a 50nm polycarbonate film (Whatman), and the number of extrusions is 10-15 times. After extrusion, use 0.22 A ⁇ m polyethersulfone filter membrane is used to filter the liposome solution to obtain a liposome solution.
  • DLS dynamic light scattering
  • PDI polydispersity index
  • the morphology of the obtained Utenidron liposomes was characterized using a cryo-transmission electron microscope (Talos-F200C). As shown in Figure 5, the liposomes were in the form of capsules with a particle size of less than 100 nm.
  • the Utendelon liposome composition of the present invention has a smaller particle size, uniform distribution, and normality; the encapsulation efficiency is as high as more than 90%; Stored at 8°C for up to two months, the drug content is still close to 100%, and the particle size and particle size distribution have no significant changes; it has a sustained release effect and prolongs the action time; the drug loading capacity is high, up to 3%. Even as high as 7%, which is far beyond the drug loading capacity of the existing utideron preparations, so that the amount of excipients can be reduced, the toxicity related to excipients can be reduced, and the clinical needs can be more easily met.

Landscapes

  • Health & Medical Sciences (AREA)
  • Chemical & Material Sciences (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Medicinal Chemistry (AREA)
  • Pharmacology & Pharmacy (AREA)
  • Animal Behavior & Ethology (AREA)
  • General Health & Medical Sciences (AREA)
  • Public Health (AREA)
  • Veterinary Medicine (AREA)
  • Epidemiology (AREA)
  • Dispersion Chemistry (AREA)
  • Chemical Kinetics & Catalysis (AREA)
  • General Chemical & Material Sciences (AREA)
  • Engineering & Computer Science (AREA)
  • Oil, Petroleum & Natural Gas (AREA)
  • Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
  • Organic Chemistry (AREA)
  • Biophysics (AREA)
  • Molecular Biology (AREA)
  • Medicinal Preparation (AREA)
  • Pharmaceuticals Containing Other Organic And Inorganic Compounds (AREA)
  • Medicines That Contain Protein Lipid Enzymes And Other Medicines (AREA)

Abstract

提供了优替德隆脂质体组合物，该脂质体组合物主要包含优替德隆、磷脂以及任选的胆固醇。所述脂质体组合物不含有易引起人体过敏反应的辅料，具有较高的载药量和稳定性，具有良好的产业化潜力。还提供了该优替德隆脂质体组合物的制备方法和用途。

Description

优替德隆脂质体组合物及其制备方法和用途 技术领域
本发明涉及医药技术领域，更具体而言涉及含优替德隆(或其盐)、磷脂以及任选的甾醇的脂质体组合物及其制备方法和用途。
背景技术
优替德隆是一类埃博霉素类衍生物，为大环内脂类化合物，由经基因修饰后的黏细菌纤维堆囊菌产生的次级代谢物。经研究表明埃博霉素类抗生素具有与紫杉醇相同的药理作用机制，其通过抑制微管蛋白的解聚而发挥抗肿瘤作用。优替德隆的化学名称为：4,8-二羟基-5,5,7,9,13-五甲基-16-[1-甲基-2-(2-甲基-噻唑-4-基)-乙烯基]-十六烷氧杂环-13-烯-2,6-酮内酯。
优替德隆易溶于乙醇、甲醇、乙酸乙酯、三氯甲烷，难溶于水，在水中的饱和溶解度低于1μg/ml，临床使用的制剂处方中含有大量的聚氧乙烯(35)蓖麻油和有机溶剂，由此具有引发严重的过敏反应的可能性，临床应用前均需给予病人进行预防性抗过敏治疗，患者的用药依从性较差。
US20090191264A和CN100409846C专利中记载将埃博霉素B制备成脂质体，但是US20090191264A说明书中记载的实施例中所使用的是天然磷脂酰胆碱或不添加胆固醇的处方，由于处方的原因，其在体内体外的稳定性可能不足以实现缓释的功能；而在CN100409846C说明书中记载埃博霉素B重量比仅为0.1％-2％。在EP2286795A1专利中记载将埃博霉素制备成阳离子脂质体，但是阳离子脂质直接应用于人体存在具有较大副作用的劣势。
优替德隆与上述文献记载中的埃博霉素类衍生物的化学结构均不相同，而药物化学结构的改变常使其理化性质发生显著的变化，从而影响药物与辅料间的相互作用关系，使优替德隆制备成脂质体组合物具有很大的困难，如本申请对比例1所示，所制备的优替德隆脂质体的稳定性较差，在制备过程中或制备完成后药液很快发生沉淀，大部分药物难以通过除菌过滤器，载药量非常低，不具有工艺可行性。
发明内容
因此，本发明所要解决的技术问题之一为提供一种稳定的优替德隆脂质体。
进一步地，本发明所要解决的技术问题是提供一种载药量高的稳定的优替德隆脂质体。
进一步地，本发明所要解决的技术问题是提供一种制备稳定的优替德隆脂质体的方法，以高效率将优替德隆稳定地包封在脂质体中。
为了解决上述技术问题，发明人创造性地运用不同类别磷脂的特殊理化特性，选择适合优替德隆的磷脂或磷脂组合，制备得到了一种或多种稳定的优替德隆脂质体。
一方面，本发明提供了一种脂质体组合物，其中所述脂质体组合物包含优替德隆和磷脂。本发明还提供了一种脂质体组合物，其中所述脂质体组合物包含优替德隆、磷脂和甾醇。在本发明的脂质体组合物中，优替德隆被包载在所述脂质体中。
在一个或多个实施方式中，所述磷脂选自聚乙二醇化磷脂、阴离子磷脂、阳离子磷脂和两性离子磷脂中的一种或多种。
在一个或多个实施方式中，所述磷脂包含两性离子磷脂或由两性离子磷脂组成。
在一个或多个实施方式中，所述磷脂包含两性离子磷脂与聚乙二醇化磷脂、阴离子磷脂和/或阳离子磷脂的组合或由所述组合组成。
在一个或多个实施方式中，所述磷脂包含两性离子磷脂与聚乙二醇化磷脂的组合或由所述组合组成。在一个或多个实施方式中，所述磷脂包含两性离子磷脂与阴离子磷脂的组合或由所述组合组成。在一个或多个实施方式中，所述磷脂包含两性离子磷脂与阳离子磷脂的组合或由所述组合组成。在一个或多个实施方式中，所述磷脂包含两性离子磷脂与聚乙二醇化磷脂和阴离子磷脂的组合或由所述组合组成。在一个或多个实施方式中，所述磷脂包含两性离子磷脂与聚乙二醇化磷脂和阳离子磷脂的组合或由所述组合组成。
在一个或多个实施方式中，所述两性离子磷脂选自卵磷脂酰胆碱(EPC)、卵磷脂酰丝氨酸(EPS)、磷脂酰乙醇胺(EPE)、大豆磷脂酰丝氨酸(SPS)、大豆磷脂酰乙醇胺(SPE)、氢化卵磷脂酰胆碱(HEPC)、氢化卵磷脂酰丝氨酸(HEPS)、氢化卵磷脂酰乙醇胺(HEPE)、氢化大豆磷脂酰胆碱 (HSPC)、氢化大豆磷脂酰丝氨酸(HSPS)、氢化大豆磷脂酰乙醇胺(HSPE)、二棕榈酰磷脂酰胆碱(DPPC)、1-棕榈酰-2-肉豆蔻酰磷脂酰胆碱(PMPC)、1-肉豆蔻酰-2-棕榈酰磷脂酰胆碱(MPPC)、二油酰磷脂酰胆碱(DOPC)、二肉豆蔻酰磷脂酰胆碱(DMPC)、二硬脂酰磷脂酰胆碱(DSPC)、1-棕榈酰-2-硬脂酰磷脂酰胆碱(PSPC)、1,2-二花生酰-sn-甘油基-3-磷脂酰胆碱(DBPC)、1-硬脂酰-2-棕榈酰磷脂酰胆碱(SPPC)、1,2-二十二碳烯酰-sn-甘油基-3-磷酸胆碱(DEPC)、棕榈酰油酰磷脂酰胆碱(POPC)、二月桂酰磷脂酰胆碱(DLPC)、棕榈酰硬脂酰磷脂酰胆碱(PSPC)、溶血磷脂酰胆碱(LPC)、二亚油酰基磷脂酰胆碱(DLPC)、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)、二油酰磷脂酰乙醇胺(dioleylphosphatidylethanolamine，DOPE)、棕榈酰油酰磷脂酰乙醇胺(POPE)中的一种或多种。
在一个或多个优选实施方式中，所述两性离子磷脂选自不饱和两性离子磷脂。不饱和两性离子磷脂的实例包括但不限于卵磷脂酰胆碱(EPC)、卵磷脂酰丝氨酸(EPS)、磷脂酰乙醇胺(EPE)、大豆磷脂酰丝氨酸(SPS)、大豆磷脂酰乙醇胺(SPE)、二油酰磷脂酰胆碱(DOPC)、1,2-二十二碳烯酰-sn-甘油基-3-磷酸胆碱(DEPC)、棕榈酰油酰磷脂酰胆碱(POPC)、二油酰磷脂酰乙醇胺(DOPE)、棕榈酰油酰磷脂酰乙醇胺(POPE)。
在一个或多个优选实施方式中，所述两性离子磷脂选自合成两性离子磷脂。合成两性离子磷脂的实例包括但不限于DSPC和DOPC。
在一个或多个更优选实施方式中，所述两性离子磷脂选自不饱和合成两性离子磷脂。所述不饱和合成两性离子磷脂的实例包括但不限于DOPC。
在一个或多个优选实施方式中，所述两性离子磷脂选自卵磷脂酰胆碱(EPC)、氢化大豆磷脂酰胆碱(HSPC)、二油酰磷脂酰胆碱(DOPC)、二硬脂酰磷脂酰胆碱(DSPC)中的一种或多种。
在一个或多个优选实施方式中，所述两性离子磷脂选自卵磷脂酰胆碱(EPC)、二油酰磷脂酰胆碱(DOPC)、二硬脂酰磷脂酰胆碱(DSPC)中的一种或多种。
在一个或多个优选实施方式中，所述两性离子磷脂选自卵磷脂酰胆碱(EPC)、二油酰磷脂酰胆碱(DOPC)中的一种或两种。
在一个或多个更优选实施方式中，所述两性离子磷脂为二油酰磷脂酰 胆碱(DOPC)。
在一个或多个实施方式中，所述聚乙二醇化磷脂选自二硬脂酰磷脂酰乙醇胺-聚乙二醇(DSPE-PEG)、二硬脂酰磷脂酰乙醇胺-甲氧基聚乙二醇(DSPE-MPEG)、二肉豆蔻酰磷脂酰乙醇胺-聚乙二醇(DMPE-PEG)、二棕榈酰甘油琥珀酸酯聚乙二醇(DPGS-PEG)、胆固醇基-聚乙二醇化磷脂和神经酰胺基聚乙二醇化磷脂中的一种或多种。
在一个或多个优选实施方式中，所述聚乙二醇化磷脂选自二硬脂酰磷脂酰乙醇胺-聚乙二醇(DSPE-PEG)和二硬脂酰磷脂酰乙醇胺-甲氧基聚乙二醇(DSPE-MPEG)中的一种或两种。
在一个或多个更优选实施方式中，所述聚乙二醇化磷脂是二硬脂酰磷脂酰乙醇胺-甲氧基聚乙二醇(DSPE-MPEG)。
在一个或多个实施方式中，所述阴离子磷脂选自二(十六烷基)磷酸酯(DhP)、磷脂酰肌醇、磷脂酰丝氨酸、磷脂酰甘油、溶血磷脂酰甘油(lysylphosphatidylglycerol，LPG)、磷脂酰乙醇胺、磷脂酸、心磷脂和胆固醇半琥珀酸酯中的一种或多种。
在一个或多个实施方式中，所述磷脂酰丝氨酸选自二肉豆蔻酰磷脂酰丝氨酸、二棕榈酰磷脂酰丝氨酸和二硬酰磷脂酰丝氨酸的一种或多种。
在一个或多个优选实施方式中，所述阴离子磷脂选自磷脂酰甘油。
在一个或多个实施方式中，所述磷脂酰甘油选自二肉豆蔻酰磷脂酰甘油(DMPG)、二棕榈酰磷脂酰甘油(DPPG)、二硬脂酰磷脂酰甘油(DSPG)、二油酰磷脂酰甘油(DOPG)、二月桂酰磷脂酰甘油(DLPG)、卵磷脂酰甘油(EPG)、卵磷脂酰肌醇(EPI)、大豆磷脂酰甘油(SPG)、大豆磷脂酰肌醇(SPI)、氢化卵磷脂酰甘油(HEPG)、氢化大豆磷脂酰甘油(HSPG)、氢化卵磷脂酰肌醇(HEPI)、和棕榈酰硬脂酰磷脂酰甘油(PSPG)、氢化大豆磷脂酰肌醇(HSPI)中的一种或多种。
在一个或多个优选实施方式中，所述磷脂酰甘油选自二硬脂酰磷脂酰甘油(DSPG)。
在一个或多个实施方式中，所述阳离子磷脂选自N-[1-(2,3-二油酰氧基)丙基]-N,N,N-三甲基铵盐(DOTAP)、N-[1-(2,3-二肉豆蔻酰基)丙基]-N,N,N-三甲基铵盐(DMTAP)、N-[1-(2,3-二棕榈酰基)丙基]-N,N,N-三甲基铵盐(DPTAP)、N-[1-(2,3-二硬脂酰基)丙基]-N,N,N-三甲基铵盐(DSTAP)、二甲 基二(十八烷基)溴化铵(DDAB)、1,2-二酰氧基-3-三甲基铵丙烷、N-[1-(2,3-二油酰氧基)丙基]-N,N-二甲基胺(DODAP)、1,2-二酰氧基-3-二甲基铵丙烷、N-[1-(2,3-二油酰氧基)丙基]-N,N,N-三甲基氯化铵(DOTMA)、1,2-二烷基氧基-3-二甲基铵丙烷、二(十八烷基)酰胺基甘氨酰基精胺(DOGS)、3-[N-(N',N'-二甲基氨基乙烷)氨基甲酰基]胆固醇(DC-Chol)、2,3-二油酰基-N-(2-(精胺酰胺)-乙基)-N,N-二甲基-1-丙铵三氟乙酸酯(DOSPA)、β-丙氨酰基胆固醇、十六烷基三甲基溴化铵(CTAB)、二C14-脒、N-叔丁基-N'十四烷基-3-十四基氨基丙酰胺(N-tert-butyl-N'-tetradecyl-3-tetradecylaminopropionamidine)、N-(α-三甲基铵乙酰基)二(十二烷基)-d-谷氨酰氯化物(TMAG)、二(十四碳酰基)-N-(三甲基铵-乙酰基)二乙醇胺氯化物、1,3-二油酰氧基-2-(6-羧基-精胺基)-丙酰胺(DOSPER)和N,N,N',N'-四甲基-N'-双(2-羟乙基)-2,3-二油酰氧基-1,4-丁烷二铵碘化物、1-[2-(酰氧基)乙基]2-烷基(链烯基)-3-(2-羟基乙基)-咪唑啉鎓氯化物衍生物如1-[2-(9(Z)-十八烯酰氧基)乙基]-2-(8(Z)-十七烯基-3-(2-羟乙基)-咪唑啉鎓氯化物(DOTIM)、1-[2-(十六烷酰氧基)乙基]-2-十五烷基-3-(2-羟乙基)咪唑啉氯化物(DPTIM)、1,2-二油酰基-3-二甲基-羟乙基溴化铵(DORI)、1,2-二油酰氧基丙基-3-二甲基-羟乙基溴化铵(DORIE)、1,2-二油酰氧基丙基-3-二甲基-羟丙基溴化铵(DORIC-HP)、1,2-二油基–氧-丙基-3-二甲基-羟乙基溴化铵(DORIE-HB)、1,2-二油酰氧基丙基-3-二甲基-羟戊基溴化铵(DORIE-Hpe)、1,2-二肉豆蔻氧基丙基-3-二甲基-羟乙基溴化铵(DMRIE)、1,2-二棕榈酰氧基丙基-3-二甲基-羟乙基溴化铵(DPRIE)、1,2-二癸氧基丙基-3-二甲基-羟乙基溴化铵(DMRIE)、1,2-二硬脂氧基-N,N-二甲基-3-氨基丙烷(DSDMA)、1,2-二油酰氧基-N,N-二甲基-3-氨基丙酸(DODMA)、1,2-二亚油基氧基-N,N-二甲基-3-氨基丙烷(DLinDMA)、1,2-二亚麻基氧基-N,N-二甲基-3-氨基丙烷(DLenDMA)中的一种或多种。
在一个或多个实施方式中，所述磷脂包含不饱和两性离子磷脂与阴性离子磷脂的组合或由所述组合组成。在一个或多个实施方式中，所述磷脂包含合成两性离子磷脂与阴性离子磷脂的组合或由所述组合组成。在一个或多个优选实施方式中，所述磷脂包含不饱和合成两性离子磷脂与阴性离子磷脂的组合或由其组成。在一个或多个更优选实施方式中，所述磷脂包含二油酰磷脂酰胆碱(DOPC)和二硬脂酰磷脂酰甘油(DSPG)的组合或由其组成。
在一个或多个实施方式中，所述磷脂包含不饱和两性离子磷脂与聚乙二醇化磷脂的组合或由所述组合组成。在一个或多个实施方式中，所述磷脂包含合成两性离子磷脂与聚乙二醇化磷脂的组合或由所述组合组成。在一个或多个优选实施方式中，所述磷脂包含不饱和合成两性离子磷脂与聚乙二醇化磷脂的组合或由所述组合组成。在一个或多个更优选实施方式中，所述磷脂包含二油酰磷脂酰胆碱(DOPC)和二硬脂酰磷脂酰乙醇胺-甲氧基聚乙二醇(DSPE-MPEG)的组合或由所述组合组成。
在一个或多个实施方式中，所述甾醇选自胆固醇、7-氢化胆固醇、羊毛固醇、谷甾醇、菜子固醇、真菌固醇、牡蛎固醇、豆甾醇、麦角固醇中的一种或多种。在一个或多个优选实施方式中，所述甾醇选自胆固醇和麦角固醇中的一种或两种。在一个或多个更优选实施方式中，所述甾醇为胆固醇。
在一个或多个实施方式中，所述脂质体组合物包含优替德隆和两性离子磷脂，例如如上所述的两性离子磷脂，优选选自EPC、DOPC和DSPC中的一种或多种。在一个或多个更优选实施方式中，所述脂质体组合物包含优替德隆和DOPC。
在一个或多个实施方式中，所述脂质体组合物包含优替德隆、两性离子磷脂和聚乙二醇化磷脂。在一个或多个实施方式中，所述脂质体组合物包含优替德隆、DSPE-MPEG和两性离子磷脂，其中所述两性离子磷脂是如上所述的两性离子磷脂，优选选自HSPC、EPC、DOPC和DSPC中的一种或多种，优选选自HSPC、EPC和DOPC中的一种或多种。
在一个或多个优选实施方式中，所述脂质体组合物包含优替德隆、两性离子磷脂和甾醇。在一个或多个更优选实施方式中，所述脂质体组合物包含优替德隆、两性离子磷脂和胆固醇，其中所述两性离子磷脂是如上所述的两性离子磷脂，优选选自HSPC、EPC、DOPC和DSPC中的一种或多种，更优选选自EPC、DOPC和DSPC中的一种或多种。在一个或多个更优选实施方式中，所述脂质体组合物包含优替德隆、EPC和胆固醇。在一个或多个更优选实施方式中，所述脂质体组合物包含优替德隆、DOPC、DSPC和胆固醇。
在一个或多个优选实施方式中，所述脂质体组合物包含优替德隆、聚乙二醇化磷脂、两性离子磷脂和甾醇。在一个或多个优选实施方式中，所 述脂质体组合物包含优替德隆、DSPE-MPEG、两性离子磷脂和胆固醇，其中所述两性离子磷脂是如上所述的两性离子磷脂，优选选自DOPC、DSPC、HSPC和EPC中的一种或多种，更优选选自DOPC、HSPC和EPC中的一种或多种。在一个或多个更优选实施方式中，所述脂质体组合物包含优替德隆、DSPE-MPEG、DOPC和胆固醇。
在一个或多个优选实施方式中，所述脂质体组合物包含优替德隆、阴离子磷脂、两性离子磷脂和甾醇。在一个或多个优选实施方式中，所述脂质体组合物包含优替德隆、DSPG、两性离子磷脂和胆固醇，其中所述两性离子磷脂是如上所述的两性离子磷脂，优选选自DOPC、DSPC、HSPC和EPC中的一种或多种，更优选选自DOPC、DSPC和EPC中的一种或多种。在一个或多个更优选实施方式中，所述脂质体组合物包含优替德隆、DSPG、DOPC和胆固醇。
在一个实施方式中，所述脂质体组合物不包括仅由优替德隆和HSPC组成的脂质体组合物。
在一个或多个实施方式中，在所述脂质体组合物中，按脂质体组合物的总重量计，优替德隆的含量为2.5-20重量％，优选为3-10重量％。
在一个或多个实施方式中，在所述脂质体组合物中，按脂质体组合物的总重量计，所述磷脂的含量为40.0-97.5重量％，优选为65.0-97.0重量％。
在一个或多个实施方式中，在所述脂质体组合物中，当含有甾醇时，按脂质体组合物的总重量计，所述甾醇的含量为0.8-48.8重量％，优选为0.9-48.5重量％，更优选3-30重量％。
在一个或多个实施方式中，在所述脂质体组合物中，当所述磷脂为两性离子磷脂与聚乙二醇化磷脂、阴离子磷脂和/或阳离子磷脂的组合时，按磷脂的总重量计，所述两性离子磷脂的含量为50-90重量％；所述聚乙二醇化磷脂的含量为0-40重量％；所述阴离子磷脂的含量为0-40重量％；所述阳离子磷脂的含量为0-40重量％。
在一个或多个实施方式中，在所述脂质体组合物中，当所述磷脂为两性离子磷脂与聚乙二醇化磷脂的组合时，按磷脂的总重量计，所述两性离子磷脂的含量为60％-90％，优选为70％-85％；所述聚乙二醇化磷脂的含量为10-40％，优选为15％-30％。
在一个或多个实施方式中，在所述脂质体组合物中，当所述磷脂为两 性离子磷脂与阴离子磷脂的组合时，按磷脂的总重量计，所述两性离子磷脂的含量为60％-90％，优选为65％-85％，更优选为70％-80％；所述阴离子磷脂的含量为10-40％，优选为15％-35％，更优选为20％-30％。
在一个或多个实施方式中，在所述脂质体组合物中，当所述磷脂为两性离子磷脂与阳离子磷脂的组合时，按磷脂的总重量计，所述两性离子磷脂的含量为60％-90％，优选为65％-85％，更优选为70％-80％；所述阳离子磷脂的含量为10-40％，优选为15％-35％，更优选为20％-30％。
在一个或多个实施方式中，优替德隆与总磷脂的质量比为0.2％至30％，优选为2.6-25.3％，更优选为2.6％-11.2％，更优选为3.0-8.0％。
在一个或多个实施方式中，当脂质体组合物中存在甾醇时，甾醇与总磷脂的质量比为1％-50％，优选为3％-40％，更优选为5％-35％。
在一个或多个实施方式中，脂质体组合物还包含渗透压调节剂、抗氧化剂、防腐剂、pH调节剂、缓冲剂中的一种或多种。在一个或多个实施方式中，所述渗透压调节剂选自氯化钠、甘油、山梨醇、甘露醇和葡萄糖中的一种或多种。在一个或多个实施方式中，所述pH调节剂选自氢氧化钠、柠檬酸钠、柠檬酸、磷酸、醋酸和盐酸中的一种或多种。在一个或多个实施方式中，所述防腐剂选自羟苯烷基酯类、苯甲酸、苯甲酸钠、山梨酸、醋酸氯乙定和苯扎溴铵中的一种或多种。在一个或多个实施方式中，所述抗氧化剂选自亚硫酸钠、亚硫酸氢钠、焦亚硫酸钠、硫代硫酸钠、抗坏血酸、叔丁基对羟基茴香醚、2,6-二叔丁基化羟基甲苯和维生素E中的一种或多种。在一个或多个实施方式中，所述缓冲剂选自柠檬酸盐缓冲液、磷酸盐缓冲液、醋酸盐缓冲液和Tris缓冲液中的一种或多种。
在一个或多个实施方式中，脂质体组合物还包含去离子水、10％蔗糖水溶液、磷酸盐缓冲液、柠檬酸盐缓冲液中的一种。
在一个或多个实施方式中，脂质体组合物为液体形式。在液体形式的脂质体组合物中优替德隆的浓度为0.01mg/ml-20mg/ml，优选为0.5mg/ml-5mg/ml。
在一个或多个实施方式中，脂质体组合物为冻干粉形式。在一个或多个实施方式中，冻干粉形式的脂质体组合物还可以含有冻干保护剂。在一个或多个实施方式中，所述冻干保护剂选自葡萄糖、蔗糖、麦芽糖、乳糖、甘露糖、海藻糖、甘氨酸、右旋糖酐中的一种或多种。
在一个或多个实施方式中，脂质体组合物中的脂质体粒径为小于250nm，优选为50nm至220nm。
在一个或多个实施方式中，脂质体组合物中的脂质体平均多分散指数(PDI)为小于0.3，优选小于0.2。
在一个或多个实施方式中，本发明的脂质体组合物可以通过选自剪切混合法、薄膜水化法、喷雾干燥法、冷冻干燥法、冻融法、溶剂注入法、逆相蒸发法、乳化挥发法、微流控法、超声法、超临界流体法、均质法中的一种或多种方法进行制备。
在一个或多个实施方式中，本发明的脂质体组合物可以通过薄膜水化法进行制备。在一个或多个实施方式中，所述薄膜水化法包括以下步骤：
(1)将优替德隆、磷脂、以及任选的甾醇与溶剂混合均匀，得到混合溶液；
(2)将步骤(1)中获得的混合溶液进行减压蒸发得到含优替德隆的脂膜；
(3)将步骤(2)中获得的脂膜进行水化得到脂质体溶液；
(4)将步骤(3)中获得的脂质体溶液进行整粒得到纳米脂质体溶液。
在一个或多个实施方式中，所述步骤(1)中使用的溶剂为有机溶剂或有机溶剂与水的混合物。在一个或多个实施方式中，所述有机溶剂选自氯仿、二氯甲烷、叔丁醇、异丙醇、乙酸乙酯、乙醇、甲醇、四氢呋喃、二氧六环、乙腈、丙酮、二甲基亚砜、二甲基甲酰胺、甲基吡咯烷酮中的一种或多种。
在一个或多个实施方式中，所述步骤(1)中使用的溶剂选自二氯甲烷与甲醇的混合物；二氯甲烷与乙醇的混合物；以及氯仿、甲醇和水的混合物。
在一个或多个实施方式中，所述步骤(1)中使用的溶剂选自二氯甲烷与甲醇5:1或6:1的混合物；二氯甲烷与乙醇5:1的混合物；以及氯仿、甲醇和水95:4:1的混合物。
在一个或多个实施方式中，所述步骤(3)中使用水化溶剂对脂膜进行水化。在一个或多个实施方式中，所述水化溶剂为去离子水、10％蔗糖水溶液、磷酸盐缓冲液、柠檬酸盐缓冲液中的一种或多种。在一个或多个实施方式中，所述水化溶剂还含有渗透压调节剂、抗氧化剂、防腐剂、pH调节剂、缓冲剂中的一种或多种。
在一个或多个实施方式中，在所述步骤(4)中，所述整粒使用选自剪切法、高压均质法和微射流均质法中的一种或多种方法进行。
在一个或多个实施方式中，所述薄膜水化法还包括步骤(5)：将步骤(4)中获得的纳米脂质体溶液进行除菌。
在一个或多个实施方式中，所述薄膜水化法还包括将步骤(5)得到的药物溶液冻干的步骤。
另一方面，本发明提供了本发明所述的脂质体组合物在制备用于预防或治疗癌症的药物中的用途。
在一个或多个实施方式中，所述癌症为实体瘤，如乳腺癌、肺癌及消化道肿瘤等。
在一个或多个实施方式中，所述脂质体组合物被制备成药物制剂。在一个或多个实施方式中，所述脂质体组合物被制备成通过非肠道、通过吸入、腹腔内、膀胱内、肌肉内、静脉内、气管内、皮下、眼内、鞘内、透皮给药、直肠或阴道内给药的药物制剂。在一个或多个优选实施方式中，所述脂质体组合物被制备成通过静脉内给药的药物制剂。在一个或多个优选实施方式中，所述药物制剂为固体制剂、液体制剂或气体制剂，更优选为注射用液体制剂。在一个或多个优选实施方式中，所述药物制剂为在使用前由所述脂质体组合物无菌冻干粉重构的稳定的水悬浮液。
在一个或多个实施方式中，由脂质体组合物制备的药物制剂还包含其它药物。在一个或多个实施方式中，所述其它药物为抗癌药物。
本发明的技术方案具有以下有益效果：
本发明提供了一种稳定的优替德隆脂质体组合物。
本发明的优替德隆脂质体组合物不含有易引起人体过敏反应的辅料，可有效消除现有临床制剂由于蓖麻油引发的过敏反应及有机溶剂引发的刺激性反应，从而增加患者的用药依从性，同时延长优替德隆的体内半衰期，达到减毒增效的临床效果。
本发明采用的配方组合可以使优替德隆在脂质体中的重量比增加至3％以上，该量的增加意味着载药量的增加，本申请脂质体的载药量可达3％以上，甚至高达7％，远远高于现有优替德隆制剂的载药量，载药量增加可以减少辅料用量，降低生产成本，降低患者用药负担，同时减轻辅料可能 带来的副作用。
本发明的优替德隆脂质体组合物具有较高的载药量和稳定性，具有良好的产业化潜力。
附图说明
图1表示实施例1制备的优替德隆脂质体的水化粒径分布图。
图2表示实施例5制备的优替德隆脂质体的体外释放度检测结果。
图3表示实施例6制备的优替德隆脂质体的水化粒径分布图。
图4表示实施例10制备的优替德隆脂质体的水化粒径分布图。
图5表示实施例17制备的优替德隆脂质体的形貌表征图。
具体实施方式
以下实施例是对本发明的进一步说明，并非本发明范围的限制。下面参考实施例进一步详细阐述本发明，但是本领域技术人员应当理解，本发明并不限于这些实施例以及使用的制备方法。而且，本领域技术人员根据本发明的描述可以对本发明进行等同替换、组合、改良或修饰，但这些都将包括在本发明的范围内。
测试例1脂质体的粒径检测
使用马尔文纳米粒径仪NANO ZS90采用动态光散射(DLS)检测脂质体的粒径,检测模式：自动；检测待测样品折射率：1.340；分散介质：超纯水；温度25℃；黏度：0.8872cp；分散介质折射率：1.330，得到粒径图。
测试例2脂质体中优替德隆的含量测定
含量测定按照高效液相色谱法(中国药典2020年版四部通则0512)进行。
色谱柱：GL Sciences Intertsil ODS-3 5μm，4.6*250mm
柱温：30℃；
运行时间：25min；
流速：1.0ml/min；
进样量：20μl；
进样盘温度：25℃；
流动相：水-乙腈(40:60)；
检测波长：250nm。
对照品溶液：取优替德隆对照品贮备液用乙腈稀释制成系列浓度，分别为：10μg/ml，20μg/ml，50μg/ml，100μg/ml，200μg/ml。
供试品溶液：取待测试样品，加入适量75％异丙醇振摇使复溶，将溶液全部转移置10ml容量瓶中，再用75％异丙醇润洗装样瓶，将润洗液转移至容量瓶中定容后，摇匀，用0.22μm有机系滤头过滤即得。
精密量取20μl，注入液相色谱仪，记录色谱图。按线性方程计算供试品溶液中优替德隆的含量。
测试例3优替德隆脂质体的稳定性测定
将过滤后的待测试脂质体溶液分装至西林瓶中，放入冻干机(Virtis AdVantage)中进行冻干，冻干参数为：-45℃预冻120分钟，升温至-30℃后一次干燥1000分钟，升温至25℃二次干燥720分钟。冻干品放入2-8℃冰箱保存，于设定时间取出冻干品，75％异丙醇复溶，检测粒径和含量。
测试例4优替德隆脂质体包封率测定
供试品溶液(包封药量)配制：取待测试的优替德隆脂质体溶液0.2ml上样于葡聚糖凝脂柱G-50柱分离，先用30ml PBS(6.8)洗脱液进行洗脱，收集前15ml洗脱液，测定其中优替德隆的含量为包封的药量。再用生理氯化钠溶液30ml洗脱剩余的游离药物。
供试品溶液(药物总量)配制：另取待测试的优替德隆脂质体溶液0.2ml，用60％乙腈溶解并稀释至10ml，摇匀，过滤，测定其中优替德隆的含量为包封和未包封的药物总量。
包封率按下式计算：
包封率(％)＝脂质体中包封的药量/药物总量×100。
测试例5优替德隆脂质体体外释放度检测
释放度检测方法如下：
1.仪器与试药：
全自动溶出仪、电子分析天平、高效液相色谱仪、pH计、优替德隆对照品、乙腈(色谱纯)、磷酸盐缓冲液(pH7.4)。
2.色谱条件：
色谱柱：Agilent ZORBAX Eclipse plus C18，4.6mm×150mm，3um
流动相A：水
流动相B：乙腈
检测波长：250nm
流速：1.0ml/min
柱温：30℃
进样量：50ul
流动相：乙腈-水(60：40)
3.溶液配制
空白溶液：pH7.4磷酸盐缓冲液：乙腈＝1：1
对照溶液：称取优替德隆对照品适量，精密称定，置容量瓶中，用空白溶液溶解并稀释至0.05μg/ml。
供试品溶液配制：精密吸取待测试的优替德隆脂质体溶液0.5ml，置于透析袋中，两段扎紧后，以500ml磷酸盐缓冲盐(pH7.4)为释放介质，转速100转/分，温度37±0.5℃，分别于0.5h、1.0h、2.0h、3.0h、4.0h、6.0h、10.0h、14.0h、18.0h、20.0h、22.0h、24.0h取释放介质。初滤液4ml，取样体积8ml，并同时补充空白释放介质12.0mL，将各取样点样品精密取1ml于烧杯中，再加入1ml乙腈混匀，过滤，处理后进样分析。系统平衡后，依次进样，记录色谱图并将试验结果记录于相关记录中。按如下公式计算累计释放度。
Figure PCTCN2022103468-appb-000001
式中：
A s：供试品溶液中优替德隆的峰面积；
C r：对照溶液的浓度μg/ml；
V：供试品溶液的稀释倍数，1000；
A r：对照溶液优替德隆的平均峰面积；
M：释放介质中投入的优替德隆总量；
V 1：释放度测试取样量，12mL；
V 2：溶出介质体积，500mL。
测试例6：优替德隆脂质体的载药量测定
按照测试例2的方法检测优替德隆脂质体中的优替德隆含量(w/v％)。
按照以下方法进行优替德隆脂质体中的磷脂和甾醇的含量测定：
检测仪器：Waters Arc HPLC(2424型ELSD)
色谱条件：
流动相A：甲醇-四氢呋喃-0.17mol/L乙酸铵水溶液(89：10：1)，流动相B：4mM乙酸铵水溶液，按流动相A：流动相B为98.5：1.5进行等度洗脱，流速1.0ml/min；色谱柱：Narochrom chromcore 120 C18(3μm，4.6×150mm)；柱温30℃；漂移管温度45℃，雾化温度36℃，气体流速40psi，增益100。
对照品溶液：精密称取磷脂或甾醇置适量，用甲醇溶解并稀释成系列线形对照溶液。
供试品溶液：取待测试的优替德隆脂质体溶液0.5ml置10ml容量瓶中，甲醇超声破乳溶解，定容至刻度线，经0.45μm的微孔滤膜过滤后,备用。
精密量取10μl，注入液相色谱仪，记录色谱图。按方程计算供试品溶液中磷脂和甾醇的含量(w/v％)。
按照如下公式计算优替德隆脂质体的载药量：
优替德隆脂质体载药量＝优替德隆含量/(优替德隆含量+磷脂总含量+甾醇含量)
对比例1
称取5.3mg优替德隆、152mg HSPC使用1.5ml有机溶剂(二氯甲烷与甲醇比例为5：1)溶解，65℃旋蒸(IKA，RV10)除去有机溶剂形成脂膜，使用20ml水化溶剂(pH6.54磷酸盐缓冲液，70℃)对脂膜进行水化，水化后使用探头超声仪(新芝)对水化溶液进行超声均质，均质完成后药液呈浑浊状态，肉眼可见明显沉淀，使用0.22μm聚醚砜滤膜对脂质体溶液进行过滤，使用动态光散射(DLS)检测脂质体水化粒径为147.3nm，多分散指数(PDI) 为0.203。包封率大于90％。
实施例1
称取12.5mg优替德隆、201mg DOPC、68mg DSPE-MPEG和34mg胆固醇加入烧瓶中，再加入4ml有机溶剂(二氯甲烷与甲醇比例为5：1)溶解，40℃旋蒸(IKA，RV10)除去有机溶剂形成脂膜，使用55-65℃的水化溶剂(含10％蔗糖的pH5.54柠檬酸盐缓冲液，6ml)对脂膜进行水化，水化后使用挤出器(Avanti)对水化溶液进行挤出，挤出膜为100nm，挤出次数为10-15次，挤出完成后使用0.22μm聚醚砜滤膜对脂质体溶液进行过滤，使用动态光散射(DLS)检测脂质体水化粒径。粒径图如图1所示，平均粒径为181.3nm，PDI为0.067。
实施例2
称取6.2mg优替德隆、82.5mg DOPC、30mg DSPG和15mg胆固醇加入烧瓶中，再加入2ml有机溶剂(三氯甲烷、甲醇与水的比例为95：4：1)溶解，40℃旋蒸(IKA，RV10)除去有机溶剂形成脂膜，使用40-45℃的水化溶剂(10％蔗糖溶液，5ml)对脂膜进行水化，水化后使用挤出器(Avanti)对水化溶液进行挤出，挤出膜为100nm，挤出次数为10-15次，挤出完成后使用0.22μm聚醚砜滤膜对脂质体溶液进行过滤，将过滤后的药液分装至西林瓶中，放入冻干机中进行冻干，冻干程序如测试例3中所述。冻干品放入2-8℃冰箱保存，于设定时间取出冻干品复溶检测粒径和含量。结果如表1所示。
表1
  平均粒径(nm) 多分散指数 含量(％)
0天 181.00 0.098 100.00
7天 175.5 0.059 95.49
14天 167 0.057 103.04
1月 181.6 0.101 101.36
2月 177.3 0.082 98.48
由表1数据可知，本发明制备得到的优替德隆脂质体具有极高的稳定 性，在2-8℃下储存长达两个月的时间内，药物含量仍接近100％，粒径和粒径分布无显著变化。
实施例3
称取9.1mg优替德隆、101mg DSPC、30mg DSPG和40mg胆固醇加入烧瓶中，再加入3ml有机溶剂(三氯甲烷、甲醇与水的比例为95：4：1)溶解，70℃旋蒸(IKA，RV10)除去有机溶剂形成脂膜，使用20ml水化溶剂(pH6.54磷酸盐缓冲液，70℃)对脂膜进行水化，水化后使用均质机(ATS)对水化溶液进行均质，均质压力为1200-1500bar，均质温度为60-70℃，均质15-25次，均质完成后使用0.22μm聚醚砜滤膜对脂质体溶液进行过滤，使用动态光散射(DLS)检测脂质体水化粒径为57.97nm，多分散指数(PDI)为0.078，优替德隆含量为0.01mg/ml。包封率大于90％。
实施例4
称取5.3mg优替德隆、122mg HSPC、32mg DSPE-MPEG加入烧瓶中，再加入2ml有机溶剂(二氯甲烷与甲醇比例为5：1)溶解，70℃旋蒸(IKA，RV10)除去有机溶剂形成脂膜，使用20ml水化溶剂(pH6.54磷酸盐缓冲液，70℃)对脂膜进行水化，水化后使用均质机(ATS)对水化溶液进行均质，均质压力为1200-1500bar，均质温度为60-70℃，均质15-25次，均质完成后使用0.22μm聚醚砜滤膜对脂质体溶液进行过滤，使用动态光散射(DLS)检测脂质体水化粒径为61.33nm，多分散指数(PDI)为0.168，优替德隆含量为0.127mg/ml。包封率大于90％。
实施例5
称取5.0mg优替德隆、119mg HSPC和32mg DSPE-MPEG，使用1.5ml有机溶剂(二氯甲烷与甲醇比例为5：1)溶解，65℃旋蒸(IKA，RV10)除去有机溶剂形成脂膜，使用70℃的水化溶剂(pH6.54磷酸盐缓冲液，20ml)对脂膜进行水化，水化后使用探头超声仪(新芝)对水化溶液进行超声均质，均质完成后使用0.22μm聚醚砜滤膜对脂质体溶液进行过滤，使用动态光散射(DLS)检测脂质体水化粒径为99.13nm，多分散指数(PDI)为0.221。对所得脂质体采用测试例5所述的程序进行优替德隆脂质体体外释放度检测，结 果如图2所示，体外实验初步表明本发明的优替德隆脂质体具备缓释功效。
实施例6
称取5.6mg优替德隆、103mg EPC和30mg胆固醇加入烧瓶中，再加入2ml有机溶剂(二氯甲烷与甲醇比例为5：1)溶解，40℃旋蒸(IKA，RV10)除去有机溶剂形成脂膜，使用50-55℃的水化溶剂(10％蔗糖溶液，3ml)对脂膜进行水化，水化后使用挤出器(Avanti)对水化溶液进行挤出，挤出膜为100nm，挤出次数为10-20次，挤出完成后使用0.22μm聚醚砜滤膜对脂质体溶液进行过滤，使用动态光散射(DLS)检测脂质体水化粒径为144.5nm，多分散指数(PDI)为0.069。粒径图如图3所示。包封率为94.3％。
实施例7
称取5mg优替德隆和105mg DOPC加入烧瓶中，再加入2ml有机溶剂(二氯甲烷与甲醇比例为5：1)溶解，40℃旋蒸(IKA，RV10)除去有机溶剂形成脂膜，使用50-55℃的水化溶剂(10％蔗糖溶液，3ml)对脂膜进行水化，水化后使用挤出器(Avanti)对水化溶液进行挤出，挤出膜为100nm，挤出次数为10-20次，挤出完成后使用0.22μm聚醚砜滤膜对脂质体溶液进行过滤，使用动态光散射(DLS)检测脂质体水化粒径为193.2nm，多分散指数(PDI)为0.143，优替德隆含量为1.624mg/ml。包封率大于90％。
实施例8
称取5.6mg优替德隆、20mg DSPE-MPEG和104mg DOPC加入烧瓶中，再加入2ml有机溶剂(二氯甲烷与甲醇比例为5：1)溶解，40℃旋蒸(IKA，RV10)除去有机溶剂形成脂膜，使用50-55℃的水化溶剂(pH6.54磷酸盐缓冲液，3ml)对脂膜进行水化，水化后使用挤出器(Avanti)对水化溶液进行挤出，挤出膜为100nm，挤出次数为10-20次，挤出完成后使用0.22μm聚醚砜滤膜对脂质体溶液进行过滤，使用动态光散射(DLS)检测脂质体水化粒径为196.9nm，多分散指数(PDI)为0.066，优替德隆含量为1.717mg/ml。包封率大于90％。
实施例9
称取5mg优替德隆、22mg DSPE-MPEG和104mg EPC加入烧瓶中，再加入2ml有机溶剂(二氯甲烷与甲醇比例为5：1)溶解，40℃旋蒸(IKA，RV10)除去有机溶剂形成脂膜，使用50-55℃的水化溶剂(pH6.54磷酸盐缓冲液，3ml)对脂膜进行水化，水化后使用挤出器(Avanti)对水化溶液进行挤出，挤出膜为100nm，挤出次数为10-20次，挤出完成后使用0.22μm聚醚砜滤膜对脂质体溶液进行过滤，使用动态光散射(DLS)检测脂质体水化粒径为155.8nm，多分散指数(PDI)为0.079。包封率大于90％。
实施例10
称取3.12mg优替德隆、53mg DOPC、17mg DSPE-MPEG和7mg胆固醇加入烧瓶中，再加入0.6ml有机溶剂(二氯甲烷与乙醇比例为5：1)溶解，40℃旋蒸(IKA，RV10)除去有机溶剂形成脂膜，使用40-50℃的水化溶剂((10％蔗糖溶液，3ml)对脂膜进行水化，水化后使用挤出器(Avanti)对水化溶液进行挤出，挤出膜为100nm，挤出次数为10-15次，挤出完成后使用0.22μm聚醚砜滤膜对脂质体溶液进行过滤，使用动态光散射(DLS)检测脂质体水化粒径为136.9nm，多分散指数(PDI)为0.101。粒径图如图4所示。包封率大于90％。
实施例11
称取6.5mg优替德隆、100mg DOPC、35mg DSPC和17mg胆固醇加入烧瓶中，再加入2ml有机溶剂(二氯甲烷与甲醇比例为5：1)溶解，40℃旋蒸(IKA，RV10)除去有机溶剂形成脂膜，使用50-55℃的水化溶剂(pH5.54柠檬酸盐缓冲液，3ml)对脂膜进行水化，水化后使用挤出器(Avanti)对水化溶液进行挤出，挤出膜为100nm，挤出次数为10-15次，挤出完成后使用0.22μm聚醚砜滤膜对脂质体溶液进行过滤，使用动态光散射(DLS)检测脂质体水化粒径为201.9nm，多分散指数(PDI)为0.188，优替德隆含量为2.061mg/ml，包封率大于90％。
实施例12
称取3.3mg优替德隆、52mg DOPC、17mg DSPG和5mg胆固醇加入烧瓶中，再加入2ml有机溶剂(三氯甲烷、甲醇与水的比例为95：4：1)溶 解，40℃旋蒸(IKA，RV10)除去有机溶剂形成脂膜，使用35-40℃的水化溶剂(10％蔗糖溶液，3ml)对脂膜进行水化，水化后使用挤出器(Avanti)对水化溶液进行挤出，挤出膜为100nm，挤出次数为10-15次，挤出完成后使用0.22μm聚醚砜滤膜对脂质体溶液进行过滤，使用动态光散射(DLS)检测脂质体水化粒径为144.4nm，多分散指数(PDI)为0.174。包封率大于90％。
实施例13
称取3.5mg优替德隆、53mg EPC、17mg DSPG和4mg胆固醇加入烧瓶中，再加入2ml有机溶剂(三氯甲烷、甲醇与水的比例为95：4：1)溶解，40℃旋蒸(IKA，RV10)除去有机溶剂形成脂膜，使用25-35℃的水化溶剂(pH6.54磷酸盐缓冲液，3ml)对脂膜进行水化，水化后使用挤出器(Avanti)对水化溶液进行挤出，挤出膜为100nm，挤出次数为10-15次，挤出完成后使用0.22μm聚醚砜滤膜对脂质体溶液进行过滤，使用动态光散射(DLS)检测脂质体水化粒径为147.3nm，多分散指数(PDI)为0.155。包封率大于90％。
实施例14
称取3mg优替德隆、50mg EPC、17mg DSPE-PEG和4.5mg胆固醇加入烧瓶中，再加入2ml有机溶剂(三氯甲烷、甲醇与水的比例为95：4：1)溶解，40℃旋蒸(IKA，RV10)除去有机溶剂形成脂膜，使用25-35℃的水化溶剂(10％蔗糖溶液，3ml)对脂膜进行水化，水化后使用挤出器(Avanti)对水化溶液进行挤出，挤出膜为100nm，挤出次数为10-15次，挤出完成后使用0.22μm聚醚砜滤膜对脂质体溶液进行过滤，使用动态光散射(DLS)检测脂质体水化粒径为134.3nm，多分散指数(PDI)为0.11。包封率大于90％。
实施例15
称取5.2mg优替德隆、85.2mg EPC、27.5mg DSPG和7.7mg胆固醇加入烧瓶中，再加入3ml有机溶剂(三氯甲烷、甲醇与水的比例为95：4：1)溶解，40℃旋蒸(IKA，RV10)除去有机溶剂形成脂膜，使用40-50℃的水化溶剂(10％蔗糖溶液，5ml)对脂膜进行水化，水化后使用挤出器(Avanti)对水化溶液进行挤出，挤出膜为100nm，挤出次数为10-15次，挤出完成后使 用0.22μm聚醚砜滤膜对脂质体溶液进行过滤，过滤后的药液分装至西林瓶中，放入冻干机中进行冻干，冻干参数为：-45℃预冻120分钟，升温至-30℃后一次干燥1000分钟，升温至25℃二次干燥720分钟。冻干品复溶后使用动态光散射(DLS)检测脂质体水化粒径为183.4nm，多分散指数(PDI)为0.163，优替德隆含量为0.797mg/ml。包封率大于90％。
实施例16
称取5mg优替德隆，86mg EPC，28mg DSPE-PEG和8mg胆固醇加入烧瓶中，再加入2ml有机溶剂(二氯甲烷与甲醇的比例为5：1)溶解，40℃旋蒸(IKA，RV10)除去有机溶剂形成脂膜，使用40-50℃的水化溶剂(10％蔗糖溶液，5ml)对脂膜进行水化，水化后使用挤出器(Avanti)对水化溶液进行挤出，挤出膜为100nm，挤出次数为10-15次，挤出完成后使用0.22μm聚醚砜滤膜对脂质体溶液进行过滤，过滤后的药液分装至西林瓶中，放入冻干机中进行冻干，冻干参数为：-45℃预冻120分钟，升温至-30℃后一次干燥1000分钟，升温至25℃二次干燥720分钟。冻干品复溶后使用动态光散射(DLS)检测脂质体水化粒径为173.1nm，多分散指数(PDI)为0.129，优替德隆含量为0.799mg/ml。包封率大于90％。
实施例17
称取6.7mg优替德隆、115.9mg DOPC、33.2mg DSPE-MPEG和16.7mg胆固醇加入烧瓶中，再加入2.4ml有机溶剂(二氯甲烷与乙醇比例为5：1)溶解，再加入3g玻璃珠(亚速旺，0.350-0.500mm，BZ04)，40℃旋蒸(Buchi，R300)除去有机溶剂形成脂膜，使用25-35℃的水化溶剂(纯化水，6ml)对脂膜进行水化，水化后使用挤出器(ATS，AE001)对水化溶液进行挤出，挤出膜为50nm聚碳酸酯膜(Whatman)，挤出次数为10-15次，挤出完成后使用0.22μm聚醚砜滤膜对脂质体溶液进行过滤，得到脂质体溶液。使用动态光散射(DLS)检测脂质体水化粒径为82.3nm，多分散指数(PDI)为0.075。
使用冷冻透射电镜(Talos-F200C)对获得的优替德隆脂质体进行形貌表征，如图5所示，脂质体呈囊状结构，粒径小于100nm。
使用测试例6所述的方法测定获得的优替德隆脂质体溶液中优替德隆的含量为0.84mg/ml，DOPC和DSPE-MPEG的总含量为14.55mg/ml，胆 固醇的含量为1.67mg/ml，按照所述公式进行计算，优替德隆脂质体载药量为4.9％(w/w％)。
由上述实施例可知，本发明的优替德隆脂质体组合物粒径较小，且分布均匀，呈正态性；包封率高达90％以上；具有极高的稳定性，在2-8℃下储存长达两个月的时间内，药物含量仍接近100％，粒径和粒径分布无显著变化；具备缓释作用，延长作用时间；载药量高，可达3％以上，甚至高达7％，远远超出现有优替德隆制剂的载药量，从而能够减少辅料用量，降低辅料相关毒性，更易满足临床需求。

Claims (25)

  1. 脂质体组合物，其中所述脂质体组合物包含优替德隆、磷脂以及任选的甾醇。
  2. 根据权利要求1所述的脂质体组合物，其中所述磷脂选自聚乙二醇化磷脂、阴离子磷脂、阳离子磷脂或两性离子磷脂中的一种或多种。
  3. 根据权利要求2所述的脂质体组合物，其中所述磷脂选自聚乙二醇化磷脂、阴离子磷脂或两性离子磷脂中的一种或多种；优选地，所述磷脂选自聚乙二醇化磷脂和两性离子磷脂的组合或阴离子磷脂和两性离子磷脂的组合。
  4. 根据权利要求3所述的脂质体组合物，其中所述聚乙二醇化磷脂选自二硬脂酰磷脂酰乙醇胺-聚乙二醇(DSPE-PEG)、二硬脂酰磷脂酰乙醇胺-甲氧基聚乙二醇(DSPE-MPEG)、二肉豆蔻酰磷脂酰乙醇胺-聚乙二醇(DMPE-PEG)、二棕榈酰甘油琥珀酸酯聚乙二醇(DPGS-PEG)、胆固醇基-聚乙二醇化磷脂或神经酰胺基聚乙二醇化磷脂中的一种或多种；优选选自二硬脂酰磷脂酰乙醇胺-聚乙二醇(DSPE-PEG)和二硬脂酰磷脂酰乙醇胺-甲氧基聚乙二醇(DSPE-MPEG)；更优选为二硬脂酰磷脂酰乙醇胺-甲氧基聚乙二醇(DSPE-MPEG)。
  5. 根据权利要求3所述的脂质体组合物，其中所述阴离子磷脂选自磷脂酰甘油、二(十六烷基)磷酸酯(DhP)、磷脂酰肌醇、磷脂酰丝氨酸、磷脂酰甘油、溶血磷脂酰甘油(lysylphosphatidylglycerol，LPG)、磷脂酰乙醇胺、磷脂酸、心磷脂或胆固醇半琥珀酸酯中的一种或多种；优选为磷脂酰甘油。
  6. 根据权利要求5所述的脂质体组合物，其中所述磷脂酰甘油选自二肉豆蔻酰磷脂酰甘油(DMPG)、二棕榈酰磷脂酰甘油(DPPG)、二硬脂酰磷脂酰甘油(DSPG)、二油酰磷脂酰甘油(DOPG)、二月桂酰磷脂酰甘油(DLPG)、卵磷脂酰甘油(EPG)、卵磷脂酰肌醇(EPI)、大豆磷脂酰甘油(SPG)、大豆磷脂酰肌醇(SPI)、氢化卵磷脂酰甘油(HEPG)、氢化大豆磷脂酰甘油(HSPG)、氢化卵磷脂酰肌醇(HEPI)、棕榈酰硬脂酰磷脂酰甘油(PSPG)、氢化大豆磷脂酰肌醇(HSPI)中的一种或多种；优选选自二硬脂酰磷脂酰甘油(DSPG)、氢化卵磷脂酰甘油(HEPG)、氢化大豆磷脂酰甘油(HSPG)中的一种或多种；优选为二硬脂酰磷脂酰甘油(DSPG)。
  7. 根据权利要求3所述的脂质体组合物，其中所述两性离子磷脂选自 卵磷脂酰胆碱(EPC)、卵磷脂酰丝氨酸(EPS)、磷脂酰乙醇胺(EPE)、大豆磷脂酰丝氨酸(SPS)、大豆磷脂酰乙醇胺(SPE)、氢化卵磷脂酰胆碱(HEPC)、氢化卵磷脂酰丝氨酸(HEPS)、氢化卵磷脂酰乙醇胺(HEPE)、氢化大豆磷脂酰胆碱(HSPC)、氢化大豆磷脂酰丝氨酸(HSPS)、氢化大豆磷脂酰乙醇胺(HSPE)、二棕榈酰磷脂酰胆碱(DPPC)、1-棕榈酰-2-肉豆蔻酰磷脂酰胆碱(PMPC)、1-肉豆蔻酰-2-棕榈酰磷脂酰胆碱(MPPC)、二油酰磷脂酰胆碱(DOPC)、二肉豆蔻酰磷脂酰胆碱(DMPC)、二硬脂酰磷脂酰胆碱(DSPC)、1-棕榈酰-2-硬脂酰磷脂酰胆碱(PSPC)、1,2-二花生酰-sn-甘油基-3-磷脂酰胆碱(DBPC)、1-硬脂酰-2-棕榈酰磷脂酰胆碱(SPPC)、1,2-二十二碳烯酰-sn-甘油基-3-磷酸胆碱(DEPC)、棕榈酰油酰磷脂酰胆碱(POPC)、二月桂酰磷脂酰胆碱(DLPC)、棕榈酰硬脂酰磷脂酰胆碱(PSPC)、溶血磷脂酰胆碱(LPC)、二亚油酰基磷脂酰胆碱(DLPC)、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)、二油酰磷脂酰乙醇胺(dioleyl phosphatidylethanolamine，DOPE)、棕榈酰油酰磷脂酰乙醇胺(POPE)、鞘磷脂中的一种或多种；优选选自卵磷脂酰胆碱(EPC)、氢化大豆磷脂酰胆碱(HSPC)、二油酰磷脂酰胆碱(DOPC)、二硬脂酰磷脂酰胆碱(DSPC)中的一种或多种；更优选选自卵磷脂酰胆碱(EPC)、二油酰磷脂酰胆碱(DOPC)中的一种或两种；更优选选自二油酰磷脂酰胆碱(DOPC)。
  8. 根据权利要求1-7中任一项所述的脂质体组合物，其中所述甾醇选自胆固醇、7-氢化胆固醇、羊毛固醇、谷甾醇、菜子固醇、真菌固醇、牡蛎固醇、豆甾醇、麦角固醇中的一种或多种；优选选自胆固醇。
  9. 根据权利要求1-8中任一项所述的脂质体组合物，其中当所述磷脂包含两性离子磷脂与聚乙二醇化磷脂、阴离子磷脂和/或阳离子磷脂的组合时，按磷脂的总重量计，所述两性离子磷脂的含量为50％-90％，所述聚乙二醇化磷脂的含量为0％-40％，所述阴离子磷脂的含量为0％-40％，所述阳离子磷脂的含量为0％-40％。
  10. 根据权利要求1-9中任一项所述的脂质体组合物，其中所述优替德隆与总磷脂的质量比为0.2％至30％；和/或，所述甾醇与总磷脂的质量比为0％至60％。
  11. 根据权利要求1-10中任一项所述的脂质体组合物，其中所述脂质体组合物中的脂质体直径为50nm-250nm。
  12. 根据权利要求1-11中任一项所述的脂质体组合物，其中所述脂质体组合物中的脂质体平均多分散指数小于0.25。
  13. 根据权利要求1-12中任一项所述的脂质体组合物，其中所述脂质体组合物为液体形式。
  14. 根据权利要求1-12中任一项所述的脂质体组合物，其中所述脂质体组合物为冻干粉形式。
  15. 根据权利要求13所述的脂质体组合物，其中所述脂质体组合物还含有或不含有冻干保护剂；优选地，其中所述冻干保护剂选自葡萄糖、蔗糖、麦芽糖、乳糖、甘露糖、海藻糖、甘氨酸、右旋糖酐中的一种或多种。
  16. 根据权利要求1-15中任一项所述的脂质体组合物，其还包含渗透压调节剂、抗氧化剂、防腐剂、pH调节剂、缓冲剂中的一种或多种；优选地，所述渗透压调节剂选自氯化钠、甘油、山梨醇、甘露醇和葡萄糖中的一种或多种；优选地，所述pH调节剂选自氢氧化钠、柠檬酸钠、柠檬酸、磷酸、醋酸和盐酸中的一种或多种；优选地，所述防腐剂选自羟苯烷基酯类、苯甲酸、苯甲酸钠、山梨酸、醋酸氯乙定和苯扎溴铵中的一种或多种；优选地，所述抗氧化剂选自亚硫酸钠、亚硫酸氢钠、焦亚硫酸钠、硫代硫酸钠、抗坏血酸、叔丁基对羟基茴香醚、2,6-二叔丁基化羟基甲苯和维生素E中的一种或多种；优选地，所述缓冲剂选自柠檬酸盐缓冲液、磷酸盐缓冲液、醋酸盐缓冲液和Tris缓冲液中的一种或多种。
  17. 根据权利要求1-16中任一项所述的脂质体组合物，其通过选自剪切混合法、薄膜水化法、喷雾干燥法、冷冻干燥法、冻融法、溶剂注入法、逆相蒸发法、乳化挥发法、微流控法、超声法、超临界流体法、均质法中的一种或多种方法制备得到。
  18. 制备权利要求1-16中任一项所述的脂质体组合物的方法，其为薄膜水化法，包括以下步骤：
    (1)将优替德隆、磷脂、以及任选的甾醇与溶剂混合均匀，得到混合溶液；
    (2)将步骤(1)中获得的混合溶液进行减压蒸发得到含优替德隆的脂膜；
    (3)将步骤(2)中获得的脂膜进行水化得到脂质体溶液；
    (4)将步骤(3)中获得的脂质体溶液进行整粒得到纳米脂质体溶液；
    (5)将步骤(4)中获得的纳米脂质体溶液进行除菌。
  19. 根据权利要求18所述的方法，其中步骤(1)使用的溶剂为有机溶剂或有机溶剂与水的混合物，优选所述有机溶剂选自氯仿、二氯甲烷、叔丁醇、异丙醇、乙酸乙酯、乙醇、甲醇、四氢呋喃、二氧六环、乙腈、丙酮、二甲基亚砜、二甲基甲酰胺、甲基吡咯烷酮中的一种或多种。
  20. 根据权利要求18所述的方法，其中步骤(3)所述水化所使用的溶液选自含渗透压调节剂、抗氧化剂、防腐剂、pH调节剂和缓冲剂中的一种或多种的溶液。
  21. 根据权利要求18所述的方法，其中步骤(4)所述整粒使用选自剪切法、高压均质法和微射流均质法中的一种或多种方法进行。
  22. 根据权利要求18所述的方法，其还包括将步骤(5)得到的药物溶液冻干的步骤。
  23. 权利要求1-16中任一项所述的脂质体组合物在制备用于预防或治疗癌症的药物中的用途。
  24. 药物制剂，其包含权利要求1-16中任一项所述的脂质体组合物，优选所述药物制剂为通过非肠道、通过吸入、腹腔内、膀胱内、肌肉内、静脉内、气管内、皮下、眼内，鞘内、透皮给药、直肠或阴道内给药的药物制剂，优选所述药物制剂为通过静脉内给药的药物制剂；优选所述药物制剂为固体制剂、液体制剂或气体制剂，更优选为注射用液体制剂；更进一步优选所述药物制剂为由所述脂质体组合物的无菌冻干粉重构的稳定的水悬浮液。
  25. 根据权利要求24所述的药物制剂，其还包含其它药物，优选所述其它药物为抗癌药物。
PCT/CN2022/103468 2021-12-31 2022-07-01 优替德隆脂质体组合物及其制备方法和用途 Ceased WO2023123986A1 (zh)

Priority Applications (4)

Application Number Priority Date Filing Date Title
US18/264,506 US20240108579A1 (en) 2021-12-31 2022-07-01 Utidelone liposome composition, and preparation method therefor and use thereof
CN202280010753.3A CN116710085A (zh) 2021-12-31 2022-07-01 优替德隆脂质体组合物及其制备方法和用途
EP22913275.8A EP4275676B1 (en) 2021-12-31 2022-07-01 Utidelone liposome composition, and preparation method therefor and use thereof
JP2023542005A JP2024505154A (ja) 2021-12-31 2022-07-01 ウチデロンリポソーム組成物、及びその製造方法並びに使用

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
CN202111672446 2021-12-31
CN202111672446.5 2021-12-31

Publications (1)

Publication Number Publication Date
WO2023123986A1 true WO2023123986A1 (zh) 2023-07-06

Family

ID=86997375

Family Applications (1)

Application Number Title Priority Date Filing Date
PCT/CN2022/103468 Ceased WO2023123986A1 (zh) 2021-12-31 2022-07-01 优替德隆脂质体组合物及其制备方法和用途

Country Status (5)

Country Link
US (1) US20240108579A1 (zh)
EP (1) EP4275676B1 (zh)
JP (1) JP2024505154A (zh)
CN (1) CN116710085A (zh)
WO (1) WO2023123986A1 (zh)

Cited By (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN117224483A (zh) * 2023-09-01 2023-12-15 山东京卫制药有限公司 一种适用于热辅助治疗的硝酸2-(4-甲基噻唑-5-基)乙酯盐脑靶向脂质体

Families Citing this family (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN117731661A (zh) * 2023-11-30 2024-03-22 上海市第一人民医院 优替德隆在胰腺癌的应用

Citations (7)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN1813721A (zh) * 2005-11-22 2006-08-09 菏泽睿鹰制药集团有限公司 一种埃坡霉素b脂质体制剂及应用
US20090191264A1 (en) 1999-08-04 2009-07-30 Jean-Claude Sonntag Epothilone compositions
EP2286795A1 (en) 2002-06-26 2011-02-23 MediGene AG Method of producing a cationic liposomal preparation comprising a lipophilic compound
CN109863165A (zh) * 2016-08-23 2019-06-07 爱姆维恩公司 新型免疫刺激肽
WO2021180111A1 (zh) * 2020-03-10 2021-09-16 正大天晴药业集团股份有限公司 包括吡啶并[1,2-a]嘧啶酮化合物的药物组合
CN113402509A (zh) * 2021-06-17 2021-09-17 山东大学 一组美沙达唑类化合物及其制备方法和应用
WO2021204188A1 (zh) * 2020-04-08 2021-10-14 北京华昊中天生物医药股份有限公司 优替德隆半水合物单晶及其制备方法与应用

Family Cites Families (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
AU2001266583A1 (en) * 2000-05-26 2001-12-11 Kosan Biosciences, Inc. Epothilone derivatives and methods for making and using the same
DE60330407D1 (de) * 2002-08-23 2010-01-14 Sloan Kettering Inst Cancer Synthese von Epothilonen, Zwischenprodukte davon, Analoga und ihre Verwendung
EA200501252A1 (ru) * 2003-02-03 2006-02-24 Неофарм, Инк. Липосомальная композиция, способ ее получения и лекарственное средство на ее основе (варианты)
US10369104B2 (en) * 2014-04-01 2019-08-06 Children's Hospital Los Angeles Targeted polymerized nanoparticles for cancer treatment

Patent Citations (8)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US20090191264A1 (en) 1999-08-04 2009-07-30 Jean-Claude Sonntag Epothilone compositions
EP2286795A1 (en) 2002-06-26 2011-02-23 MediGene AG Method of producing a cationic liposomal preparation comprising a lipophilic compound
CN1813721A (zh) * 2005-11-22 2006-08-09 菏泽睿鹰制药集团有限公司 一种埃坡霉素b脂质体制剂及应用
CN100409846C (zh) 2005-11-22 2008-08-13 菏泽睿鹰制药集团有限公司 一种埃坡霉素b脂质体制剂及应用
CN109863165A (zh) * 2016-08-23 2019-06-07 爱姆维恩公司 新型免疫刺激肽
WO2021180111A1 (zh) * 2020-03-10 2021-09-16 正大天晴药业集团股份有限公司 包括吡啶并[1,2-a]嘧啶酮化合物的药物组合
WO2021204188A1 (zh) * 2020-04-08 2021-10-14 北京华昊中天生物医药股份有限公司 优替德隆半水合物单晶及其制备方法与应用
CN113402509A (zh) * 2021-06-17 2021-09-17 山东大学 一组美沙达唑类化合物及其制备方法和应用

Non-Patent Citations (1)

* Cited by examiner, † Cited by third party
Title
See also references of EP4275676A4

Cited By (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN117224483A (zh) * 2023-09-01 2023-12-15 山东京卫制药有限公司 一种适用于热辅助治疗的硝酸2-(4-甲基噻唑-5-基)乙酯盐脑靶向脂质体
CN117224483B (zh) * 2023-09-01 2024-05-10 山东京卫制药有限公司 一种适用于热辅助治疗的硝酸2-(4-甲基噻唑-5-基)乙酯盐脑靶向脂质体

Also Published As

Publication number Publication date
EP4275676A1 (en) 2023-11-15
JP2024505154A (ja) 2024-02-05
EP4275676C0 (en) 2025-08-13
CN116710085A (zh) 2023-09-05
EP4275676B1 (en) 2025-08-13
EP4275676A4 (en) 2024-07-03
US20240108579A1 (en) 2024-04-04

Similar Documents

Publication Publication Date Title
US10022365B2 (en) Liposome of irinotecan or irinotecan hydrochloride and preparation method thereof
RU2641605C2 (ru) Улучшенные липосомальные составы липофильных соединений
US5858397A (en) Liposomal formulations of mitoxantrone
EP2680820B1 (en) Advanced active liposomal loading of poorly water-soluble substances
US9655848B2 (en) Liposomes for in-vivo delivery
WO2019209787A1 (en) Inhalable liposomal sustained release composition for use in treating pulmonary diseases
US20090041835A1 (en) Method of inhibiting leakage of drug encapsulated in liposomes
DE60122304T2 (de) Auf lipiden basierendes system zur zielgerichteten verabreichung diagnostischer wirkstoffe
AU2003270102B2 (en) Non-vesicular cationic lipid formulations
Li et al. Encapsulation of mitoxantrone into pegylated SUVs enhances its antineoplastic efficacy
WO1999030686A1 (en) Cationic drugs encapsulated in anionic liposomes
WO2023123986A1 (zh) 优替德隆脂质体组合物及其制备方法和用途
US20120100067A1 (en) Solubilisation Method
CA2508166A1 (en) Compositions and methods related to lipid:emodin formulations
CN112654345A (zh) 缓释眼用药物组合物及其用途
US20130189352A1 (en) Liposome comprising combination of chloroquine and adriamycin and preparation method thereof
EP1547580A1 (en) Loading of a camptothecin drug into colloidal nanoparticles
WO2022050369A1 (ja) 抗菌薬を含有するリポソーム製剤
JPWO2005123078A1 (ja) 水難溶性カンプトテシン含有リポソーム製剤
WO2000009071A2 (en) A novel liposomal formulation useful in treatment of cancer and other proliferation diseases
JP2007262026A (ja) リポソームの製造方法
HK40046575A (zh) 緩釋眼用藥物組合物及其用途
JPWO2001000173A1 (ja) リポソームに内包された薬物の漏出抑制方法

Legal Events

Date Code Title Description
WWE Wipo information: entry into national phase

Ref document number: 2023542005

Country of ref document: JP

WWE Wipo information: entry into national phase

Ref document number: 202280010753.3

Country of ref document: CN

WWE Wipo information: entry into national phase

Ref document number: 18264506

Country of ref document: US

121 Ep: the epo has been informed by wipo that ep was designated in this application

Ref document number: 22913275

Country of ref document: EP

Kind code of ref document: A1

ENP Entry into the national phase

Ref document number: 2022913275

Country of ref document: EP

Effective date: 20230808

NENP Non-entry into the national phase

Ref country code: DE

WWG Wipo information: grant in national office

Ref document number: 2022913275

Country of ref document: EP