CN108404142A - 一种磁共振成像纳米载体、纳米载药系统及其制备方法 - Google Patents

一种磁共振成像纳米载体、纳米载药系统及其制备方法 Download PDF

Info

Publication number
CN108404142A
CN108404142A CN201810117789.7A CN201810117789A CN108404142A CN 108404142 A CN108404142 A CN 108404142A CN 201810117789 A CN201810117789 A CN 201810117789A CN 108404142 A CN108404142 A CN 108404142A
Authority
CN
China
Prior art keywords
plga
nano
dox
magnetic resonance
spio
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.)
Granted
Application number
CN201810117789.7A
Other languages
English (en)
Other versions
CN108404142B (zh
Inventor
陈填烽
罗良平
史长征
高鹏
陈樑
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.)
Jinan University
University of Jinan
Original Assignee
Jinan University
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 Jinan University filed Critical Jinan University
Priority to CN201810117789.7A priority Critical patent/CN108404142B/zh
Publication of CN108404142A publication Critical patent/CN108404142A/zh
Application granted granted Critical
Publication of CN108404142B publication Critical patent/CN108404142B/zh
Active legal-status Critical Current
Anticipated expiration legal-status Critical

Links

Classifications

    • AHUMAN NECESSITIES
    • A61MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61KPREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K49/00Preparations for testing in vivo
    • A61K49/06Nuclear magnetic resonance [NMR] contrast preparations; Magnetic resonance imaging [MRI] contrast preparations
    • A61K49/08Nuclear magnetic resonance [NMR] contrast preparations; Magnetic resonance imaging [MRI] contrast preparations characterised by the carrier
    • A61K49/10Organic compounds
    • A61K49/12Macromolecular compounds
    • A61K49/126Linear polymers, e.g. dextran, inulin, PEG
    • AHUMAN NECESSITIES
    • A61MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61KPREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K31/00Medicinal preparations containing organic active ingredients
    • A61K31/70Carbohydrates; Sugars; Derivatives thereof
    • A61K31/7028Compounds having saccharide radicals attached to non-saccharide compounds by glycosidic linkages
    • A61K31/7034Compounds having saccharide radicals attached to non-saccharide compounds by glycosidic linkages attached to a carbocyclic compound, e.g. phloridzin
    • A61K31/704Compounds having saccharide radicals attached to non-saccharide compounds by glycosidic linkages attached to a carbocyclic compound, e.g. phloridzin attached to a condensed carbocyclic ring system, e.g. sennosides, thiocolchicosides, escin, daunorubicin
    • AHUMAN NECESSITIES
    • A61MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61KPREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K47/00Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient
    • A61K47/50Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates
    • A61K47/51Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the non-active ingredient being a modifying agent
    • A61K47/54Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the non-active ingredient being a modifying agent the modifying agent being an organic compound
    • A61K47/545Heterocyclic compounds
    • AHUMAN NECESSITIES
    • A61MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61KPREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K47/00Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient
    • A61K47/50Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates
    • A61K47/51Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the non-active ingredient being a modifying agent
    • A61K47/62Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the non-active ingredient being a modifying agent the modifying agent being a protein, peptide or polyamino acid
    • A61K47/64Drug-peptide, drug-protein or drug-polyamino acid conjugates, i.e. the modifying agent being a peptide, protein or polyamino acid which is covalently bonded or complexed to a therapeutically active agent
    • AHUMAN NECESSITIES
    • A61MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61KPREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K47/00Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient
    • A61K47/50Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates
    • A61K47/69Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the conjugate being characterised by physical or galenical forms, e.g. emulsion, particle, inclusion complex, stent or kit
    • A61K47/6921Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the conjugate being characterised by physical or galenical forms, e.g. emulsion, particle, inclusion complex, stent or kit the form being a particulate, a powder, an adsorbate, a bead or a sphere
    • A61K47/6927Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the conjugate being characterised by physical or galenical forms, e.g. emulsion, particle, inclusion complex, stent or kit the form being a particulate, a powder, an adsorbate, a bead or a sphere the form being a solid microparticle having no hollow or gas-filled cores
    • A61K47/6929Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the conjugate being characterised by physical or galenical forms, e.g. emulsion, particle, inclusion complex, stent or kit the form being a particulate, a powder, an adsorbate, a bead or a sphere the form being a solid microparticle having no hollow or gas-filled cores the form being a nanoparticle, e.g. an immuno-nanoparticle
    • A61K47/6931Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the conjugate being characterised by physical or galenical forms, e.g. emulsion, particle, inclusion complex, stent or kit the form being a particulate, a powder, an adsorbate, a bead or a sphere the form being a solid microparticle having no hollow or gas-filled cores the form being a nanoparticle, e.g. an immuno-nanoparticle the material constituting the nanoparticle being a polymer
    • A61K47/6935Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the conjugate being characterised by physical or galenical forms, e.g. emulsion, particle, inclusion complex, stent or kit the form being a particulate, a powder, an adsorbate, a bead or a sphere the form being a solid microparticle having no hollow or gas-filled cores the form being a nanoparticle, e.g. an immuno-nanoparticle the material constituting the nanoparticle being a polymer the polymer being obtained otherwise than by reactions involving carbon to carbon unsaturated bonds, e.g. polyesters, polyamides or polyglycerol
    • A61K47/6937Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the conjugate being characterised by physical or galenical forms, e.g. emulsion, particle, inclusion complex, stent or kit the form being a particulate, a powder, an adsorbate, a bead or a sphere the form being a solid microparticle having no hollow or gas-filled cores the form being a nanoparticle, e.g. an immuno-nanoparticle the material constituting the nanoparticle being a polymer the polymer being obtained otherwise than by reactions involving carbon to carbon unsaturated bonds, e.g. polyesters, polyamides or polyglycerol the polymer being PLGA, PLA or polyglycolic acid
    • AHUMAN NECESSITIES
    • A61MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61KPREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K47/00Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient
    • A61K47/50Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates
    • A61K47/69Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the conjugate being characterised by physical or galenical forms, e.g. emulsion, particle, inclusion complex, stent or kit
    • A61K47/6921Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the conjugate being characterised by physical or galenical forms, e.g. emulsion, particle, inclusion complex, stent or kit the form being a particulate, a powder, an adsorbate, a bead or a sphere
    • A61K47/6927Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the conjugate being characterised by physical or galenical forms, e.g. emulsion, particle, inclusion complex, stent or kit the form being a particulate, a powder, an adsorbate, a bead or a sphere the form being a solid microparticle having no hollow or gas-filled cores
    • A61K47/6929Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the conjugate being characterised by physical or galenical forms, e.g. emulsion, particle, inclusion complex, stent or kit the form being a particulate, a powder, an adsorbate, a bead or a sphere the form being a solid microparticle having no hollow or gas-filled cores the form being a nanoparticle, e.g. an immuno-nanoparticle
    • A61K47/6931Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the conjugate being characterised by physical or galenical forms, e.g. emulsion, particle, inclusion complex, stent or kit the form being a particulate, a powder, an adsorbate, a bead or a sphere the form being a solid microparticle having no hollow or gas-filled cores the form being a nanoparticle, e.g. an immuno-nanoparticle the material constituting the nanoparticle being a polymer
    • A61K47/6939Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the conjugate being characterised by physical or galenical forms, e.g. emulsion, particle, inclusion complex, stent or kit the form being a particulate, a powder, an adsorbate, a bead or a sphere the form being a solid microparticle having no hollow or gas-filled cores the form being a nanoparticle, e.g. an immuno-nanoparticle the material constituting the nanoparticle being a polymer the polymer being a polysaccharide, e.g. starch, chitosan, chitin, cellulose or pectin
    • AHUMAN NECESSITIES
    • A61MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61KPREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K49/00Preparations for testing in vivo
    • A61K49/06Nuclear magnetic resonance [NMR] contrast preparations; Magnetic resonance imaging [MRI] contrast preparations
    • A61K49/08Nuclear magnetic resonance [NMR] contrast preparations; Magnetic resonance imaging [MRI] contrast preparations characterised by the carrier
    • A61K49/10Organic compounds
    • AHUMAN NECESSITIES
    • A61MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61KPREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K49/00Preparations for testing in vivo
    • A61K49/06Nuclear magnetic resonance [NMR] contrast preparations; Magnetic resonance imaging [MRI] contrast preparations
    • A61K49/08Nuclear magnetic resonance [NMR] contrast preparations; Magnetic resonance imaging [MRI] contrast preparations characterised by the carrier
    • A61K49/10Organic compounds
    • A61K49/14Peptides, e.g. proteins
    • AHUMAN NECESSITIES
    • A61MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61KPREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K49/00Preparations for testing in vivo
    • A61K49/06Nuclear magnetic resonance [NMR] contrast preparations; Magnetic resonance imaging [MRI] contrast preparations
    • A61K49/18Nuclear magnetic resonance [NMR] contrast preparations; Magnetic resonance imaging [MRI] contrast preparations characterised by a special physical form, e.g. emulsions, microcapsules, liposomes
    • A61K49/1818Nuclear magnetic resonance [NMR] contrast preparations; Magnetic resonance imaging [MRI] contrast preparations characterised by a special physical form, e.g. emulsions, microcapsules, liposomes particles, e.g. uncoated or non-functionalised microparticles or nanoparticles
    • A61K49/1821Nuclear magnetic resonance [NMR] contrast preparations; Magnetic resonance imaging [MRI] contrast preparations characterised by a special physical form, e.g. emulsions, microcapsules, liposomes particles, e.g. uncoated or non-functionalised microparticles or nanoparticles coated or functionalised microparticles or nanoparticles
    • A61K49/1824Nuclear magnetic resonance [NMR] contrast preparations; Magnetic resonance imaging [MRI] contrast preparations characterised by a special physical form, e.g. emulsions, microcapsules, liposomes particles, e.g. uncoated or non-functionalised microparticles or nanoparticles coated or functionalised microparticles or nanoparticles coated or functionalised nanoparticles
    • A61K49/1827Nuclear magnetic resonance [NMR] contrast preparations; Magnetic resonance imaging [MRI] contrast preparations characterised by a special physical form, e.g. emulsions, microcapsules, liposomes particles, e.g. uncoated or non-functionalised microparticles or nanoparticles coated or functionalised microparticles or nanoparticles coated or functionalised nanoparticles having a (super)(para)magnetic core, being a solid MRI-active material, e.g. magnetite, or composed of a plurality of MRI-active, organic agents, e.g. Gd-chelates, or nuclei, e.g. Eu3+, encapsulated or entrapped in the core of the coated or functionalised nanoparticle
    • A61K49/1833Nuclear magnetic resonance [NMR] contrast preparations; Magnetic resonance imaging [MRI] contrast preparations characterised by a special physical form, e.g. emulsions, microcapsules, liposomes particles, e.g. uncoated or non-functionalised microparticles or nanoparticles coated or functionalised microparticles or nanoparticles coated or functionalised nanoparticles having a (super)(para)magnetic core, being a solid MRI-active material, e.g. magnetite, or composed of a plurality of MRI-active, organic agents, e.g. Gd-chelates, or nuclei, e.g. Eu3+, encapsulated or entrapped in the core of the coated or functionalised nanoparticle having a (super)(para)magnetic core coated or functionalised with a small organic molecule
    • AHUMAN NECESSITIES
    • A61MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61KPREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K49/00Preparations for testing in vivo
    • A61K49/06Nuclear magnetic resonance [NMR] contrast preparations; Magnetic resonance imaging [MRI] contrast preparations
    • A61K49/18Nuclear magnetic resonance [NMR] contrast preparations; Magnetic resonance imaging [MRI] contrast preparations characterised by a special physical form, e.g. emulsions, microcapsules, liposomes
    • A61K49/1818Nuclear magnetic resonance [NMR] contrast preparations; Magnetic resonance imaging [MRI] contrast preparations characterised by a special physical form, e.g. emulsions, microcapsules, liposomes particles, e.g. uncoated or non-functionalised microparticles or nanoparticles
    • A61K49/1821Nuclear magnetic resonance [NMR] contrast preparations; Magnetic resonance imaging [MRI] contrast preparations characterised by a special physical form, e.g. emulsions, microcapsules, liposomes particles, e.g. uncoated or non-functionalised microparticles or nanoparticles coated or functionalised microparticles or nanoparticles
    • A61K49/1824Nuclear magnetic resonance [NMR] contrast preparations; Magnetic resonance imaging [MRI] contrast preparations characterised by a special physical form, e.g. emulsions, microcapsules, liposomes particles, e.g. uncoated or non-functionalised microparticles or nanoparticles coated or functionalised microparticles or nanoparticles coated or functionalised nanoparticles
    • A61K49/1827Nuclear magnetic resonance [NMR] contrast preparations; Magnetic resonance imaging [MRI] contrast preparations characterised by a special physical form, e.g. emulsions, microcapsules, liposomes particles, e.g. uncoated or non-functionalised microparticles or nanoparticles coated or functionalised microparticles or nanoparticles coated or functionalised nanoparticles having a (super)(para)magnetic core, being a solid MRI-active material, e.g. magnetite, or composed of a plurality of MRI-active, organic agents, e.g. Gd-chelates, or nuclei, e.g. Eu3+, encapsulated or entrapped in the core of the coated or functionalised nanoparticle
    • A61K49/1851Nuclear magnetic resonance [NMR] contrast preparations; Magnetic resonance imaging [MRI] contrast preparations characterised by a special physical form, e.g. emulsions, microcapsules, liposomes particles, e.g. uncoated or non-functionalised microparticles or nanoparticles coated or functionalised microparticles or nanoparticles coated or functionalised nanoparticles having a (super)(para)magnetic core, being a solid MRI-active material, e.g. magnetite, or composed of a plurality of MRI-active, organic agents, e.g. Gd-chelates, or nuclei, e.g. Eu3+, encapsulated or entrapped in the core of the coated or functionalised nanoparticle having a (super)(para)magnetic core coated or functionalised with an organic macromolecular compound, i.e. oligomeric, polymeric, dendrimeric organic molecule
    • A61K49/1857Nuclear magnetic resonance [NMR] contrast preparations; Magnetic resonance imaging [MRI] contrast preparations characterised by a special physical form, e.g. emulsions, microcapsules, liposomes particles, e.g. uncoated or non-functionalised microparticles or nanoparticles coated or functionalised microparticles or nanoparticles coated or functionalised nanoparticles having a (super)(para)magnetic core, being a solid MRI-active material, e.g. magnetite, or composed of a plurality of MRI-active, organic agents, e.g. Gd-chelates, or nuclei, e.g. Eu3+, encapsulated or entrapped in the core of the coated or functionalised nanoparticle having a (super)(para)magnetic core coated or functionalised with an organic macromolecular compound, i.e. oligomeric, polymeric, dendrimeric organic molecule the organic macromolecular compound being obtained otherwise than by reactions only involving carbon-to-carbon unsaturated bonds, e.g. PLGA
    • AHUMAN NECESSITIES
    • A61MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61KPREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K49/00Preparations for testing in vivo
    • A61K49/06Nuclear magnetic resonance [NMR] contrast preparations; Magnetic resonance imaging [MRI] contrast preparations
    • A61K49/18Nuclear magnetic resonance [NMR] contrast preparations; Magnetic resonance imaging [MRI] contrast preparations characterised by a special physical form, e.g. emulsions, microcapsules, liposomes
    • A61K49/1818Nuclear magnetic resonance [NMR] contrast preparations; Magnetic resonance imaging [MRI] contrast preparations characterised by a special physical form, e.g. emulsions, microcapsules, liposomes particles, e.g. uncoated or non-functionalised microparticles or nanoparticles
    • A61K49/1821Nuclear magnetic resonance [NMR] contrast preparations; Magnetic resonance imaging [MRI] contrast preparations characterised by a special physical form, e.g. emulsions, microcapsules, liposomes particles, e.g. uncoated or non-functionalised microparticles or nanoparticles coated or functionalised microparticles or nanoparticles
    • A61K49/1824Nuclear magnetic resonance [NMR] contrast preparations; Magnetic resonance imaging [MRI] contrast preparations characterised by a special physical form, e.g. emulsions, microcapsules, liposomes particles, e.g. uncoated or non-functionalised microparticles or nanoparticles coated or functionalised microparticles or nanoparticles coated or functionalised nanoparticles
    • A61K49/1827Nuclear magnetic resonance [NMR] contrast preparations; Magnetic resonance imaging [MRI] contrast preparations characterised by a special physical form, e.g. emulsions, microcapsules, liposomes particles, e.g. uncoated or non-functionalised microparticles or nanoparticles coated or functionalised microparticles or nanoparticles coated or functionalised nanoparticles having a (super)(para)magnetic core, being a solid MRI-active material, e.g. magnetite, or composed of a plurality of MRI-active, organic agents, e.g. Gd-chelates, or nuclei, e.g. Eu3+, encapsulated or entrapped in the core of the coated or functionalised nanoparticle
    • A61K49/1851Nuclear magnetic resonance [NMR] contrast preparations; Magnetic resonance imaging [MRI] contrast preparations characterised by a special physical form, e.g. emulsions, microcapsules, liposomes particles, e.g. uncoated or non-functionalised microparticles or nanoparticles coated or functionalised microparticles or nanoparticles coated or functionalised nanoparticles having a (super)(para)magnetic core, being a solid MRI-active material, e.g. magnetite, or composed of a plurality of MRI-active, organic agents, e.g. Gd-chelates, or nuclei, e.g. Eu3+, encapsulated or entrapped in the core of the coated or functionalised nanoparticle having a (super)(para)magnetic core coated or functionalised with an organic macromolecular compound, i.e. oligomeric, polymeric, dendrimeric organic molecule
    • A61K49/1863Nuclear magnetic resonance [NMR] contrast preparations; Magnetic resonance imaging [MRI] contrast preparations characterised by a special physical form, e.g. emulsions, microcapsules, liposomes particles, e.g. uncoated or non-functionalised microparticles or nanoparticles coated or functionalised microparticles or nanoparticles coated or functionalised nanoparticles having a (super)(para)magnetic core, being a solid MRI-active material, e.g. magnetite, or composed of a plurality of MRI-active, organic agents, e.g. Gd-chelates, or nuclei, e.g. Eu3+, encapsulated or entrapped in the core of the coated or functionalised nanoparticle having a (super)(para)magnetic core coated or functionalised with an organic macromolecular compound, i.e. oligomeric, polymeric, dendrimeric organic molecule the organic macromolecular compound being a polysaccharide or derivative thereof, e.g. chitosan, chitin, cellulose, pectin, starch
    • AHUMAN NECESSITIES
    • A61MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61KPREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K49/00Preparations for testing in vivo
    • A61K49/06Nuclear magnetic resonance [NMR] contrast preparations; Magnetic resonance imaging [MRI] contrast preparations
    • A61K49/18Nuclear magnetic resonance [NMR] contrast preparations; Magnetic resonance imaging [MRI] contrast preparations characterised by a special physical form, e.g. emulsions, microcapsules, liposomes
    • A61K49/1818Nuclear magnetic resonance [NMR] contrast preparations; Magnetic resonance imaging [MRI] contrast preparations characterised by a special physical form, e.g. emulsions, microcapsules, liposomes particles, e.g. uncoated or non-functionalised microparticles or nanoparticles
    • A61K49/1821Nuclear magnetic resonance [NMR] contrast preparations; Magnetic resonance imaging [MRI] contrast preparations characterised by a special physical form, e.g. emulsions, microcapsules, liposomes particles, e.g. uncoated or non-functionalised microparticles or nanoparticles coated or functionalised microparticles or nanoparticles
    • A61K49/1824Nuclear magnetic resonance [NMR] contrast preparations; Magnetic resonance imaging [MRI] contrast preparations characterised by a special physical form, e.g. emulsions, microcapsules, liposomes particles, e.g. uncoated or non-functionalised microparticles or nanoparticles coated or functionalised microparticles or nanoparticles coated or functionalised nanoparticles
    • A61K49/1827Nuclear magnetic resonance [NMR] contrast preparations; Magnetic resonance imaging [MRI] contrast preparations characterised by a special physical form, e.g. emulsions, microcapsules, liposomes particles, e.g. uncoated or non-functionalised microparticles or nanoparticles coated or functionalised microparticles or nanoparticles coated or functionalised nanoparticles having a (super)(para)magnetic core, being a solid MRI-active material, e.g. magnetite, or composed of a plurality of MRI-active, organic agents, e.g. Gd-chelates, or nuclei, e.g. Eu3+, encapsulated or entrapped in the core of the coated or functionalised nanoparticle
    • A61K49/1866Nuclear magnetic resonance [NMR] contrast preparations; Magnetic resonance imaging [MRI] contrast preparations characterised by a special physical form, e.g. emulsions, microcapsules, liposomes particles, e.g. uncoated or non-functionalised microparticles or nanoparticles coated or functionalised microparticles or nanoparticles coated or functionalised nanoparticles having a (super)(para)magnetic core, being a solid MRI-active material, e.g. magnetite, or composed of a plurality of MRI-active, organic agents, e.g. Gd-chelates, or nuclei, e.g. Eu3+, encapsulated or entrapped in the core of the coated or functionalised nanoparticle the nanoparticle having a (super)(para)magnetic core coated or functionalised with a peptide, e.g. protein, polyamino acid
    • AHUMAN NECESSITIES
    • A61MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61KPREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K9/00Medicinal preparations characterised by special physical form
    • A61K9/14Particulate form, e.g. powders, Processes for size reducing of pure drugs or the resulting products, Pure drug nanoparticles
    • A61K9/141Intimate drug-carrier mixtures characterised by the carrier, e.g. ordered mixtures, adsorbates, solid solutions, eutectica, co-dried, co-solubilised, co-kneaded, co-milled, co-ground products, co-precipitates, co-evaporates, co-extrudates, co-melts; Drug nanoparticles with adsorbed surface modifiers
    • A61K9/145Intimate drug-carrier mixtures characterised by the carrier, e.g. ordered mixtures, adsorbates, solid solutions, eutectica, co-dried, co-solubilised, co-kneaded, co-milled, co-ground products, co-precipitates, co-evaporates, co-extrudates, co-melts; Drug nanoparticles with adsorbed surface modifiers with organic compounds
    • AHUMAN NECESSITIES
    • A61MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61KPREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K9/00Medicinal preparations characterised by special physical form
    • A61K9/14Particulate form, e.g. powders, Processes for size reducing of pure drugs or the resulting products, Pure drug nanoparticles
    • A61K9/141Intimate drug-carrier mixtures characterised by the carrier, e.g. ordered mixtures, adsorbates, solid solutions, eutectica, co-dried, co-solubilised, co-kneaded, co-milled, co-ground products, co-precipitates, co-evaporates, co-extrudates, co-melts; Drug nanoparticles with adsorbed surface modifiers
    • A61K9/146Intimate drug-carrier mixtures characterised by the carrier, e.g. ordered mixtures, adsorbates, solid solutions, eutectica, co-dried, co-solubilised, co-kneaded, co-milled, co-ground products, co-precipitates, co-evaporates, co-extrudates, co-melts; Drug nanoparticles with adsorbed surface modifiers with organic macromolecular compounds
    • AHUMAN NECESSITIES
    • A61MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61PSPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P35/00Antineoplastic agents
    • BPERFORMING OPERATIONS; TRANSPORTING
    • B82NANOTECHNOLOGY
    • B82YSPECIFIC USES OR APPLICATIONS OF NANOSTRUCTURES; MEASUREMENT OR ANALYSIS OF NANOSTRUCTURES; MANUFACTURE OR TREATMENT OF NANOSTRUCTURES
    • B82Y15/00Nanotechnology for interacting, sensing or actuating, e.g. quantum dots as markers in protein assays or molecular motors
    • BPERFORMING OPERATIONS; TRANSPORTING
    • B82NANOTECHNOLOGY
    • B82YSPECIFIC USES OR APPLICATIONS OF NANOSTRUCTURES; MEASUREMENT OR ANALYSIS OF NANOSTRUCTURES; MANUFACTURE OR TREATMENT OF NANOSTRUCTURES
    • B82Y5/00Nanobiotechnology or nanomedicine, e.g. protein engineering or drug delivery

Landscapes

  • Health & Medical Sciences (AREA)
  • Chemical & Material Sciences (AREA)
  • Engineering & Computer Science (AREA)
  • Nanotechnology (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • General Health & Medical Sciences (AREA)
  • Veterinary Medicine (AREA)
  • Public Health (AREA)
  • Animal Behavior & Ethology (AREA)
  • Epidemiology (AREA)
  • Medicinal Chemistry (AREA)
  • Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
  • Bioinformatics & Cheminformatics (AREA)
  • Pharmacology & Pharmacy (AREA)
  • Radiology & Medical Imaging (AREA)
  • Molecular Biology (AREA)
  • Immunology (AREA)
  • Proteomics, Peptides & Aminoacids (AREA)
  • Crystallography & Structural Chemistry (AREA)
  • Biophysics (AREA)
  • Chemical Kinetics & Catalysis (AREA)
  • General Chemical & Material Sciences (AREA)
  • Medical Informatics (AREA)
  • Organic Chemistry (AREA)
  • General Engineering & Computer Science (AREA)
  • Biotechnology (AREA)
  • Medicinal Preparation (AREA)
  • Pharmaceuticals Containing Other Organic And Inorganic Compounds (AREA)
  • Medicines Containing Antibodies Or Antigens For Use As Internal Diagnostic Agents (AREA)
  • Medicines That Contain Protein Lipid Enzymes And Other Medicines (AREA)

Abstract

本发明属于生物医药领域,公开了一种磁共振成像纳米载体、纳米载药系统及其制备方法。本发明使用叶酸和穿膜肽两种靶向分子,提高对肿瘤的靶向效果,同时提高体内外抗肿瘤活性。并使用低毒、生物相容性良好的PLGA为抗肿瘤药物载体,并在PLGA末端修饰CS,实现功能化纳米体系的制备。同时高效负载核磁成像药物和抗肿瘤药物,使抗肿瘤药物专一到达肿瘤病灶部位,实现了四氧化三铁纳米粒子的肿瘤区域核磁定位,同时实现高选择性、低毒性的治疗效果。本发明有效克服传统细胞毒药物的选择性差、毒副作用较强、易产生耐药性等缺点,提高抗肿瘤药物的利用度和降低毒副性。载药系统制备方法简单易行,且制备的产物重复性和稳定性良好。

Description

一种磁共振成像纳米载体、纳米载药系统及其制备方法
技术领域
本发明属于生物医药领域,特别涉及一种磁共振成像纳米载体、纳米载药系统及其制备方法。
背景技术
随着发病率和死亡率的上升,癌症是导致死亡的重要原因并且成为世界性的重大公共卫生问题。然而,传统化疗药物具有非选择性,往往毒性较大,并且容易产生肿瘤的多重耐药性。新型多功能纳米载药系统是目前最具应用前景的药载传递,主要为该纳米系统能够有效提高药物生物利用度,增加药物的稳定性,延长血浆的循环时间,降低药物毒副作用以及克服肿瘤耐药性等特点,同时诊断成像并动态监测药物治疗的反应。
纳米递药系统可以通过高通透效应和高滞留效应(Enhanced permeability andretention effect,EPR effect)对肿瘤进行被动靶向作用实现在肿瘤部分的有效富集[Jhaveri AM,et al,Frontiers in Pharmacology,2014,5.]。同时可以对纳米材料进行表面修饰,将具有肿瘤靶向性的单克隆抗体配体或者特异性小分子与纳米材料相结合,利用受体与配体的特异性亲和力,构建成功能化纳米载药系统的靶向性的分子探针,显著提高肿瘤细胞对纳米药物的摄取效率,从而达到对肿瘤的主动靶作用[Yang X,et al,Biomaterials,2010,31(34):9065-9073]。
叶酸是一种小分子量的必须维生素,人体自身不能合成,故需要从外界获取并转运至细胞内。叶酸受体又被称为叶酸结合蛋白,叶酸受体(FR-α)在人体正常组织细胞表达高度保守,而在部分上皮来源的恶性肿瘤肿瘤细胞表面表达水平较高[Campbell I G,etal,Cancer Research,1991,51(19):5329-5338.]。叶酸结合的纳米载体包括脂质体、蛋白质、聚合物等,聚合物纳米由于具有表面易修饰、稳定性良好、载药量高等特点,应用前景越来越广泛。同时叶酸还具有体积小、价格低廉、对受体高度特异的亲合性、高度的化学及生物稳定性等优点。叶酸在肿瘤特异性显像及靶向治疗中的应用已成为当前研究的热点之一。
细胞穿膜肽(Cell penetrating peptides,CPPs)是近年来发现的拥有超强细胞膜穿透能力的跨膜转运分子,具有低裂解性、水溶性,并可以通过非受体依赖的内吞方式进入各种细胞膜等特点[Jewell C M,et al,Biomacromolecules,2007,8(3):857-863.]。通过改良CPPs靶细胞摄取效率低、缺乏细胞的特异性等缺点,在CPPs基础上设计合成肿瘤靶向性CPPs,又称为可活化CPPs(穿膜肽,Activatable cellpenetrating peptide,ACPP,氨基酸序列为E8-PLGLAG-R9-C)。穿膜肽(ACPP)是一种具有肿瘤微环境响应的多肽,一般穿膜肽(ACPP)包括了A、B、C三段序列的多肽,能靶向MMP-2和MMP-9。A段是活性中心(CPP区),由寡聚精氨酸构成,未被封闭时有穿膜活性,B段序列是识别位点,而C段序列带负电荷,通过中和A段的正电荷从而封闭其穿膜活性,而当A段与C段之间的链接区域被蛋白酶裂解时,释放出A段从而发挥穿膜功能,从而促进纳米粒子进入肿瘤细胞[Huang S,et al,Biomaterials,2013,34(21):5294-5302.]。目前穿膜肽(ACPP)作为新型的药物载体工具,已经成为研究热点。
自从1999年Weissldeer[Weissleder R.Radiology,1999,212(3):609-614.]提出分子影像(Molecular imaging,MI)的概念以来,分子影像学技术迅猛发展,其在肿瘤早期诊断、活体成像、治疗检测以及疗效评价等方面发挥着重要作用。Fe3O4纳米粒子(SPIO)是一种磁共振阴性造影剂,具有优异的超顺磁性能。超顺磁性是指纳米颗粒粒径在10nm到20nm之间或者更小时具有单畴结构的铁磁物质,在室温下表现为顺磁性特点,但在外磁场作用下其顺磁性磁化率远高于一般顺磁材料的磁化率,称为超顺磁性。超顺磁性SPIO纳米粒子的主要作用是改变核磁共振的R2驰豫,缩短T2时间,减弱T2加权信号。通过静脉注射将超顺磁性SPIO纳米粒子注入到体内后,它会和血浆蛋白结合被枯否氏细胞吞噬,除了集中分布在肝、脾、淋巴结等具有丰富网状内皮细胞的组织和器官,肿瘤组织也有丰富的网状内皮细胞,可以富集磁性纳米粒子,因此随着具有更长血液半衰期的SPIO的出现,SPIO作为磁性造影剂能够达到肿瘤细胞、分子水平的显像,提高核磁共振成像检测技术的灵敏度。
在此基础上,联合并交叉多学科的不同优势,通过优化材料构建高效、稳定及安全的纳米体系平台,并利用该平台装载高效的抗肿瘤药物、高灵敏度的肿瘤诊断探针和高亲和力的肿瘤靶向基元,组成集合靶向肿瘤药物运输、释放、治疗以及预后监测等功能于一体的多功能化的纳米体系平台,将成为具有创新性和应用前景的研究领域。
发明内容
为了克服上述现有技术的缺点与不足,本发明的首要目的在于提供一种磁共振成像纳米载体。
本发明另一目的在于提供上述磁共振成像纳米载体在制备抗肿瘤药物中的应用。
本发明另一目的在于提供一种磁共振成像纳米载药系统。
本发明另一目的在于提供上述磁共振成像纳米载药系统的制备方法。
本发明的目的通过下述方案实现:
一种磁共振成像纳米载体,其为聚合物纳米粒子,所述的聚合物为PLGA-CS,其生物相容性良好,且其表面具有活性基团。
所述的聚合物纳米粒子的尺寸为100~300纳米;
所述的活性基团为氨基、羟基或羧基。
本发明的磁共振成像纳米药物载体的原料为聚乳酸-羟基乙酸共聚物(PLGA)和壳聚糖(CS),具有低毒,且具有良好的生物相容性的优点,是抗肿瘤药物载体的理想来源;将其制成尺寸为100~300纳米的纳米粒子,不仅可以通过静电或氢键等相互作用高效负载各类抗肿瘤药物,具有普遍适用性,而且由于纳米粒子表面具有亲水性基团,可以显著提高癌细胞对载药聚合物纳米粒子的摄取量,且利于发挥药物的被动靶向作用,提高局部组织的药物浓度,亲水性基团还有利于对其进行表面修饰或表面功能化,如在聚合物纳米粒子表面接合靶向分子,使其具有主动靶向能力。而本发明的纳米载体PLGA-CS,表面的活性基团包括氨基、羧基、羟基,其中氨基、羟基等亲水性基团的存在可以显著提高上述的被动靶向作用,氨基、羧基、羟基等功能基团可以为修饰基团或者功能化基团,表面修饰或表面功能化后负载抗肿瘤药物的聚合物纳米粒子可以具备主动靶向能力。
上述的磁共振成像纳米药物载体在制备抗肿瘤药物中的应用,优选在制备抗肿瘤载药系统中的应用。
所述的肿瘤包括肝癌、肺癌、恶性黑色瘤、乳腺癌、结肠癌、鼻咽癌、膀胱癌、宫颈癌、胃癌、食管癌、前列腺癌和结肠癌。
一种磁共振成像纳米载药系统,其载体即为上述的磁共振成像纳米药物载体,该载药系统主要由以下方法制备得到:
(1)将PLGA、核磁成像造影剂、抗肿瘤药物加入丙酮溶液中,得到PLGA丙酮溶液;
(2)将步骤(1)中得到的PLGA丙酮溶液逐滴加入吐温水溶液中,搅拌,得到PLGA水溶液;
(3)向步骤(2)中的得到的PLGA水溶液中加入N-羟基琥珀酰亚胺(NHS)和1-(3-二甲氨基丙基)-3-乙基碳二亚胺盐酸盐(EDC)活化PLGA上的羧基,再加入CS水溶液,搅拌,得到PLGA-CS溶液;
(4)向步骤(3)中得到的PLGA-CS溶液中加入靶向分子,搅拌,得到PLGA-CS聚合物纳米粒子水溶液,即形成磁共振成像纳米载药系统。
步骤(1)中所述的抗肿瘤药物为柔红霉素、阿霉素、去甲氧柔红霉素、表阿霉素、紫杉醇、香菇多糖、长春花碱、长春新碱、三苯氧胺、福美司坦、阿那曲唑、氟他胺、5-氟尿嘧啶、甲氨蝶呤、顺铂、卡铂、奥沙利铂、卡莫司汀、托瑞米芬、替加氟、姜黄素、去甲氧基姜黄素、双曲甲氧基姜黄素和塞替派中的至少一种;
优选的,步骤(1)中所述的抗肿瘤药物优选为阿霉素(DOX),阿霉素根据自身性质,通过亲水作用进入聚合物纳米粒子亲水性的壳中,实现聚合物载体对抗肿瘤药物的负载。功能上,阿霉素是一种抗肿瘤抗生素,可抑制RNA和DNA的合成,对RNA的抑制作用最强,从而使细胞的组成发生变异,影响细胞分裂,致使细胞死亡。因此,此类抗生素药物可高效抑制肿瘤生长,PLGA-CS聚合物纳米粒子与此类抗生素药物结合可发挥其良好抗肿瘤活性。。
步骤(1)中所述的核磁成像造影剂为超小超顺磁性氧化铁纳米粒子(SPIO)或钆喷酸葡胺注射液。
优选的,步骤(1)中所述的核磁成像造影剂为超小超顺磁性氧化铁纳米粒子(SPIO),超小超顺磁性氧化铁纳米粒子根据自身脂溶性质,通过疏水作用进入PLGA-CS聚合物纳米粒子亲脂性的核壳内,实现聚合物载体对核磁成像药物的负载。功能上,超顺磁性氧化铁(SPIO)是目前最灵敏的MR造影剂之一,其可显著缩短T2弛豫时间,使得T2加权图像变暗,因此,称为T2阴性对比剂。
步骤(1)中所得到的PLGA丙酮溶液中,PLGA的浓度为1~10mg/mL、核磁成像造影剂的浓度为1~20mg/mL、抗肿瘤药物的浓度为10~500μM。
步骤(2)中所述的逐滴是指滴入速度为每滴间隔为1~10秒,优选为5秒。
步骤(2)中所述的吐温水溶液是指浓度为1~5mg/mL的吐温水溶液,优选为5mg/mL;所述的吐温优选为吐温-80;
步骤(2)中所述的搅拌的条件优选为200~800r/min搅拌过夜。
步骤(2)中所述的PLGA水溶液中抗肿瘤药物的浓度为4~200μM。
步骤(3)中所加入的N-羟基琥珀酰亚胺(NHS)与步骤(3)中PLGA水溶液中PLGA的摩尔比为1~3:1;步骤(3)中所加入的1-(3-二甲氨基丙基)-3-乙基碳二亚胺盐酸盐(EDC)与步骤(3)中PLGA水溶液中PLGA的摩尔比为1~3:1;
步骤(3)中所述的活化PLGA上的羧基是指活化时间为2~12h;优选为4h;
步骤(3)中所述的CS水溶液的浓度为0.6~1mg/mL,优选为0.8mg/mL;所述的CS的分子量为5000~20000,所加入的CS水溶液的用量满足步骤(3)中PLGA水溶液中PLGA与CS水溶液中CS的摩尔比为1:1~10;
步骤(3)中所述的搅拌是指搅拌速度为200~800r/min,搅拌时间为8~24h。
步骤(4)中所述的靶向分子为环形RGD多肽(cRGD)、叶酸(FA)、整合素、转铁蛋白、可活化细胞穿膜肽(ACPP)、MUC-1附膜蛋白、半乳糖胺、新生血管靶向肽、粒细胞巨噬细胞刺激因子中的两种;优选为叶酸和可活化细胞穿膜肽;更优选为质量比为16:25的叶酸和可活化细胞穿膜肽。
步骤(4)中所述的靶向分子的添加量满足靶向分子与步骤(3)中加入的CS水溶液中CS的质量比为1:10。
步骤(4)中所述的搅拌是指在200~800r/min下搅拌过夜。
上述的磁共振成像纳米载药系统的制备方法中,在步骤(4)后面还包括一个纯化步骤,所述的纯化包括离心、重悬。
所述的离心是指8000~20000rpm离心5~30分钟,离心的次数为2~5次;优选的,所述的离心是指在5000rpm离心10分钟,离心次数为5次。
所述的重悬优选为用二次蒸馏水重悬。
所述的PLGA-CS聚合物纳米粒子的保存方式为:1~30℃下以溶胶或粉末形态保存。
上述步骤(1)~(4)中,若未指出温度,则均为常温条件(15~35℃)。
本发明相对于现有技术,具有如下的优点及有益效果:
1、本发明提供一种磁共振成像纳米载体、纳米载药系统及其制备方法,解决肿瘤诊断及在化学治疗过程中局部用药浓度低、全身毒性反应剧烈等问题;克服传统细胞毒药物的选择性差、毒副作用较强、易产生耐药性等缺点。本发明制备方法简单易行,且制备的产物可在水溶液中稳定保存,利于储藏。
2、本发明基于正常细胞和肿瘤细胞之间的差异,使用叶酸(FA)和穿膜肽(ACPP)两种靶向分子,赋予其更有效的靶向肿瘤的能力。并使用低毒、生物相容性良好的聚乳酸-羟基乙酸共聚物(PLGA)作为抗肿瘤药物载体,并在PLGA末端修饰壳聚糖(CS),同时高效负载核磁成像药物和抗肿瘤药物,使抗肿瘤药物专一到达肿瘤病灶部位,实现了超顺磁性四氧化三铁纳米粒子在肿瘤区域的核磁定位,提高了治疗效果的同时降低了毒副作用,实现高效低毒的治疗目标。
3、本发明的优点在于作为核磁成像抗肿瘤药物载体的聚合物纳米粒子在水溶液中稳定性好、方便储存、生物相容性好,同时,粒子表面存在各种功能基团,有利于对其进行表面修饰或表面功能化,如在聚合物纳米粒子表面接合靶向分子,使其具有主动靶向能力。
4、本发明中采用了新型材料超顺磁性四氧化三铁(SPIO),由于SPIO具有超顺磁性,更有利于药物的体内核磁成像。
5、本发明中PLGA-CS由于其CS分子具有高的正电荷,所以具有更好的稳定性,更有利于药物进入肿瘤细胞,并且CS带有大量羟基和氨基,便于纳米粒子的表面修饰。利用该载体(PLGA-CS)负载核磁成像药物和抗肿瘤药物,获得的磁共振成像纳米载药体系可实现对肿瘤的同步诊断与治疗。
附图说明
图1是实施例1中聚合物纳米粒子F/A-PLGA@DOX/SPIO的组成结构图;
图2是实施例1中聚合物纳米粒子F/A-PLGA@DOX/SPIO的红外谱图;
图3是实施例1中聚合物纳米粒子F/A-PLGA@DOX/SPIO的透射电镜图;
图4是实施例1中聚合物纳米粒子F/A-PLGA@DOX/SPIO的粒径分布;
图5是实施例1中聚合物纳米粒子F/A-PLGA@DOX/SPIO的电位分布;
图6是实施例1中聚合物纳米粒子F/A-PLGA@DOX/SPIO与单独SPIO在不同铁浓度下的1/T2值;
图7是实施例2中不同药物对不同细胞的的半数抑制浓度IC50
图8是实施例2中不同药物处理后人肺癌细胞内ROS水平的变化图;
图9是实施例2中不同药物孵育的人肺癌细胞的细胞周期分布图;
图10是实施例2中F/A-PLGA@DOX/SPIO和单独阿霉素在人肺癌细胞的细胞吸收情况图;
图11是实施例2中F/A-PLGA@DOX/SPIO和单独阿霉素在人正常肝细胞的细胞吸收情况图;
图12是人肺癌细胞,人黑色素瘤细胞,人宫颈癌细胞,人正常肝细胞对叶酸的蛋白表达图;
图13为聚合物纳米粒子F/A-PLGA@DOX/SPIO在人肺癌细胞中的荧光细胞定位图;
图14为实施例3中不同药物在接种人肺癌细胞的荷瘤裸鼠中的体内MRI定位图;
图15为实施例3不同药物在接种人肺癌细胞的荷瘤裸鼠中的体内T2的变化百分比图;
图16是实施例3中不同药物在经尾静脉注射72h后荷瘤裸鼠心、肝、脾、肺、肾和肿瘤的普鲁士蓝染色图;
图17为实施例3中聚合物纳米粒子F/A-PLGA@DOX/SPIO在大鼠中阿霉素的血药浓度图;
图18为实施例4中聚合物纳米粒子F/A-PLGA@DOX/SPIO治疗接种人肺癌细胞的荷瘤裸鼠的肿瘤照片图;
图19为实施例4中聚合物纳米粒子F/A-PLGA@DOX/SPIO治疗接种人肺癌细胞的荷瘤裸鼠的肿瘤大体标本图;
图20为实施例4中所得聚合物纳米粒子F/A-PLGA@DOX/SPIO治疗接种人肺癌细胞的荷瘤裸鼠的相对肿瘤体积图;
图21为实施例4中聚合物纳米粒子F/A-PLGA@DOX/SPIO治疗接种人肺癌细胞的荷瘤裸鼠的相对肿瘤增殖率图;
图22为实施例4中聚合物纳米粒子F/A-PLGA@DOX/SPIO治疗接种人肺癌细胞的荷瘤裸鼠的肿瘤生长抑制率图;
图23为实施例4中聚合物纳米粒子F/A-PLGA@DOX/SPIO治疗接种人肺癌细胞的荷瘤裸鼠的体重图;
图24为实施例4中聚合物纳米粒子F/A-PLGA@DOX/SPIO治疗接种人肺癌细胞的荷瘤裸鼠的T2核磁成像图;
图25为实施例4中聚合物纳米粒子F/A-PLGA@DOX/SPIO治疗接种人肺癌细胞的荷瘤裸鼠心、肝、脾、肺、肾和肿瘤的H&E染色图;
图26为实施例4中聚合物纳米粒子F/A-PLGA@DOX/SPIO治疗接种人肺癌细胞的荷瘤裸鼠的血液指标参数图;
图27为实施例4中所得聚合物纳米粒子F/A-PLGA@DOX/SPIO治疗接种人肺癌细胞的荷瘤裸鼠的H&E染色以及CD31、Ki67、VEGF免疫组化图。
具体实施方式
下面结合实施例和附图对本发明作进一步详细的描述,但本发明的实施方式不限于此。
实施例中所用试剂如无特殊说明均可从市场常规购得。A549细胞、HeLa细胞、A375细胞和L02细胞均购自于美国模式培养物保藏所,ACTT。
本发明提供了一种磁共振成像抗肿瘤药物载体,所述磁共振成像抗肿瘤药物载体为聚合物纳米粒子。所使用的原料为聚乳酸-羟基乙酸共聚物、壳聚糖。
本发明的聚合物命名中,当在PLGA末端修饰CS后,聚合物的命名中可省略CS,如实施例步骤(3)中的CS-PLGA@DOX/SPIO也可命名为PLGA@DOX/SPIO。
实施例1:聚合物纳米粒子F/A-PLGA@DOX/SPIO的制备及表征
(1)常温常压(15~35℃,1标准大气压)下,将聚乳酸-羟基乙酸共聚物(PLGA)(LA:GA=50:50,Mn=13000,美国Sigma)、超小超顺磁性氧化铁纳米粒子(SPIO)(购买于Sigma公司)和阿霉素(DOX)加入到丙酮溶液中,配置聚乳酸-羟基乙酸共聚物(PLGA)质量浓度为5mg/mL,超小超顺磁性氧化铁纳米粒子(SPIO)质量浓度为1mg/mL,阿霉素(DOX)浓度为500μM的丙酮溶液。
(2)将3mL配置好的丙酮溶液逐滴加入到10mL的吐温-80水溶液(5mg/mL)中,滴入速度为每滴间隔为5秒,200~800r/min搅拌过夜,得到阿霉素浓度为100μM的PLGA水溶液。
(3)向PLGA水溶液中加入1-(3-二甲氨基丙基)-3-乙基碳二亚胺盐酸盐(EDC)和N-羟基琥珀酰亚胺(NHS)粉末,常温下搅拌,活化PLGA上的羧基,活化时间为4小时,其中,EDC和NHS与所述PLGA水溶液中的PLGA的摩尔比都为1.5:1;再加入0.8mg/mL壳聚糖水溶液,PLGA水溶液中PLGA与CS水溶液中CS的摩尔比为1:5(CS,分子量为10000,购自于美国Sigma)反应,搅拌过夜,生成CS-PLGA@DOX/SPIO(简称为PLGA@DOX/SPIO,后面的聚合物命名中均省略CS)溶液。
(4)向生成的PLGA@DOX/SPIO溶液中加入16mg的叶酸(FA)(购买于Sigma公司)和25mg的穿膜肽(ACPP)多肽靶向分子(购买于Sigma公司),搅拌过夜,制备靶向性的聚合物纳米粒子。反应结束后,聚合物纳米粒子经过2轮离心(离心速度为5000rpm,每轮持续10分钟),再用8mL二次蒸馏水重悬,得到聚合物纳米粒子的胶体混悬液,记为F/A-PLGA@DOX/SPIO。
当将实施例1中步骤(4)中25mg的穿膜肽(ACPP,氨基酸序列为E8-PLGLAG-R9-C)多肽靶向分子替换为25mg的叶酸时得到产物记为FA-PLGA@DOX/SPIO;
当将实施例1步骤(4)中16mg的叶酸(FA)替换为16mg的穿膜肽(ACPP)多肽靶向分子时得到的产物记为ACPP-PLGA@DOX/SPIO;
当实施例1步骤(1)中不加阿霉素(DOX)时,步骤(4)得到产物记为F/A-PLGA@SPIO;
上述所得F/A-PLGA@DOX/SPIO具有较好性能,置于4℃冰箱保存。现以F/A-PLGA@DOX/SPIO为例表征其特性,具体为,F/A-PLGA@DOX/SPIO的组成结构(图1);用傅里叶变换红外光谱仪表征F/A-PLGA@DOX/SPIO的化学结构(图2);用Hitachi H-7650型透射电子显微镜表征F/A-PLGA@DOX/SPIO纳米粒子的形貌图(见图3),结果可见F/A-PLGA@DOX/SPIO纳米粒子具有良好的分散性;
用Nano-ZS(Malvern Instruments Limited)表征F/A-PLGA@DOX/SPIO水溶液的粒径(图4)及其电动电位(Zeta potential)(图5),F/A-PLGA@DOX/SPIO纳米粒子的粒径与电镜结果相符,并且具有高的表面电位,有利于肿瘤细胞的吸收;用GE1.5T Sigma HDxt磁共振成像仪表征F/A-PLGA@DOX/SPIO纳米粒子的1/T2信号(图6),结果可见F/A-PLGA@DOX/SPIO的1/T2信号跟铁浓度成剂量依赖性。
实施例2:聚合物纳米粒子F/A-PLGA@DOX/SPIO的体外抗人肺癌细胞活性研究
将A549细胞、HeLa细胞、A375细胞和L02细胞按照常规方法把细胞消化重悬后,按照肿瘤细胞密度为2×104cells/mL,正常细胞密度为4×104cells/mL,加入96孔板中,每孔100μL。培养过夜后,按照实验设置加入含有不同浓度药物(0,0.015625,0.03125,0.0625,0.125,0.25,0.5)的培养基100μL/孔,72小时后往每孔加入30μL MTT(5mg/mL),孵育3.5小时后弃去上清后再加入150μL DMSO溶解甲瓒,使用酶联免疫检测仪检测(570nm)。计算得到药物对细胞的半数抑制浓度IC50及安全系数(Safety index,SI)(图7和表1)。
表1不同药物对不同细胞的半数抑制浓度IC50及安全系数
a正常细胞。
b安全指数(SI)=IC50(正常细胞)/IC50(肿瘤细胞)。
从图7和表1可见,FA-PLGA@DOX/SPIO,ACPP-PLGA@DOX/SPIO和F/A-PLGA@DOX/SPIO对A549细胞的IC50值分别为0.095μM,0.082μM和0.069μM。其中F/A-PLGA@DOX/SPIO对A549细胞抗肿瘤活性是单独DOX的1.5倍左右。F/A-PLGA@DOX/SPIO(IC50=0.172)对L02细胞的毒性相较单独DOX(IC50=0.099)约有1.7倍左右的减低。单独DOX的SI仅为0.94,而F/A-PLGA@DOX/SPIO的SI为2.50,说明其毒性远低于DOX。
我们进一步检测F/A-PLGA@DOX/SPIO和DOX处理后A549细胞内活性氧(ROS)水平。按照常规方法把A549细胞消化重悬后,按照细胞密度为2×106cells/mL,加入96孔板中,每孔100μL,装载DHE探针,用PBS溶液洗涤后,经F/A-PLGA@DOX/SPIO和DOX作用(给药量均为1μM),立即利用荧光酶标仪持续检测DHE探针的荧光强度,同时以不加药单独细胞组作为对照组。药物处理后的人肺癌细胞内ROS水平的变化情况如图8所示,从图8中可以看出F/A-PLGA@DOX/SPIO与对照组相比能显著的提升了细胞内ROS的水平,且在0.5h内升高至最大值,约为500%。同浓度的DOX引起细胞内产生ROS量明显低于F/A-PLGA@DOX/SPIO。由于F/A-PLGA@DOX/SPIO在接上靶向分子之后,能有效地被A549细胞吸收,故使细胞产生更多的ROS。
在A549细胞和L02细胞培养24h后(4×104个细胞),分别加入不同的药物(给药量0.5μM)预处理相同时间(24h),消化收集细胞后,70%预冷乙醇处理过夜,通过PI染色后,再用流式细胞术分析经过DOX,FA-PLGA@DOX/SPIO,ACPP-PLGA@DOX/SPIO和F/A-PLGA@DOX/SPIO处理的A549细胞的细胞周期,同时以不加药单独细胞组作为对照组结果如图9所示,从图9可知,A549细胞用DOX及FA-PLGA@DOX/SPIO,ACPP-PLGA@DOX/SPIO和F/A-PLGA@DOX/SPIO处理后,细胞凋亡峰分别上升到7.8%、10.7%、20.1%和24.3%。流式细胞周期分析的结果提示F/A-PLGA@DOX/SPIO可能是通过诱导细胞凋亡的方式抑制A549细胞的增殖。
在A549细胞和L02细胞培养24h后(4×106个细胞),分别加入2μM的药物预处理不同时间,通过测定A549细胞和L02细胞内DOX浓度,结果分别如图10和11所示,从图10可见,随着时间的延长,F/A-PLGA@DOX/SPIO在A549内的细胞吸收呈现时间依赖性,在6小时达到166μg/106细胞,是单独DOX组的2.2倍。而在人正常肝细胞L02细胞中,F/A-PLGA@DOX/SPIO的细胞吸收与单独DOX组差别很小(图11)。说明F/A-PLGA@DOX/SPIO不仅明显提高了肿瘤细胞对药物的吸收效率,也使其在正常细胞和肿瘤细胞之间具有选择性。
我们又通过Western blotting检测了A549,HeLa,A375和L02细胞的叶酸的受体表达,结果如图12所示,从图12中可见,A549细胞的FR-α有较高表达,这有利于FA对A549细胞的主动靶向。接着通过细胞荧光定位实验(图13),证明了F/A-PLGA@DOX/SPIO定位于A549细胞的溶酶体。
实施例3:聚合物纳米粒子F/A-PLGA@DOX/SPIO的体内定位及药代动力学实验
制备荷瘤裸鼠模型:收集体外培养的人非小细胞肺癌细胞A549,计数,调整细胞悬液浓度为1×107个/ml,接种0.1ml细胞悬液接种于4~5周龄裸鼠(BALB/c-nu裸鼠,2~4周龄,体重约18~22g,北京华阜康生物科技有限公司)右后肢根部皮下。
分组与给药:裸鼠移植瘤用游标卡尺测量移植瘤直径,待肿瘤生长至75-100mm3后将动物随机分为三组,每组3只。A组给予SPIO(5mg/kg),B组给予F/A-PLGA@SPIO(5mg/kg),C组给予F/A-PLGA@DOX/SPIO(5mg/kg),同时开始给药,所有药物均采用尾静脉给药。同时利用核磁成像技术动态观察被试动物的核磁信号。
给药后0h、1h、4h、12h、24h、48h、72h的体内药物定位实验图如图14所示,从图14可见,F/A-PLGA@DOX/SPIO明显地影响核磁的T2成像。给药后0h、1h、4h、12h、24h、48h、72h的T2变化百分比如图15所示,从图15中可以看出在药物注射72h时F/A-PLGA@DOX/SPIO在肿瘤组织累积的量是单独SPIO的2倍以上。
实验结束后将裸鼠处死,取出全部肿瘤组织及心脏、肝脏、脾脏、肾脏及肺脏,进行普鲁士蓝染色,观测各组织内SPIO积聚情况,结果如图16所示,从图16中可以看出F/A-PLGA@SPIO、F/A-PLGA@DOX/SPIO组肿瘤组织的阳性染色率高于SPIO组,而肝、脾组织的阳性染色率明显低于SPIO组。F/A-PLGA@DOX/SPIO聚合物体系能有效地提高SPIO在肿瘤区域的累积和停留时间,有利于SPIO的体内核磁成像。
我们又通过药代动力学实验检测了F/A-PLGA@DOX/SPIO的血药浓度,结果如图17所示,从图17中可以看出,单独阿霉素组在静脉注射后,血药浓度迅速下降。而F/A-PLGA@DOX/SPIO组的血药浓度能维持较高的血药浓度,具有较高的体内循环时间,有利于药物的肿瘤治疗。
实施例4:聚合物纳米粒子F/A-PLGA@DOX/SPIO的体内抗人非小细胞肺癌细胞A549裸鼠异种移植瘤生长的抑制作用
制备荷瘤裸鼠模型:收集体外培养的人肺癌细胞A549,计数,调整细胞悬液浓度为1×107个/ml,接种0.1ml细胞悬液接种于4~5周龄裸鼠(BALB/c-nu裸鼠,2~4周龄,体重约18~22mg,北京华阜康生物科技有限公司)右后肢根部皮下。
分组与给药:裸鼠移植瘤用游标卡尺测量移植瘤直径,待肿瘤生长至75-100mm3后将动物随机分为四组,每组10只。A组给予生理盐水作为对照组;B组给予单纯DOX,浓度2mg/kg;C组给予F/A-PLGA@DOX/SPIO,浓度1mg/kg;C组给予F/A-PLGA@DOX/SPIO,浓度2mg/kg。隔天给药共持续28天,所有药物均采用尾静脉给药。使用测量瘤径的方法,动态观察被试物抗肿瘤的效应,给药28天后小鼠处死,手术剥取瘤块称重。
观测指标:
①肿瘤体积(tumor volume,TV)的计算公式为:
TV=1/2×a×b2其中a、b分别表示长宽
②根据测量的结果计算出相对肿瘤体积(relative tumor volume,RTV),计算公式为:
RTV=Vt/V0,其中V0为分笼给药时(即d0)测量所得肿瘤体积,Vt为每一次测量时的肿瘤体积。
③抗肿瘤活性的评价指标:相对肿瘤增殖率T/C(%),计算公式如下:
T/C(%)=(TRTV/CRTV)×100%,其中TRTV:治疗组RTV;CRTV:模型对照组RTV。
④抗肿瘤活性的评价指标:肿瘤生长抑制率(%),计算公式如下:
肿瘤生长抑制率(%)=100×(模型对照组平均瘤重-给药组平均瘤重/模型对照组平均瘤重)
本实验我们研究了聚合物纳米粒F/A-PLGA@DOX/SPIO治疗人肺癌细胞A549裸鼠异种移植瘤生长的抑制作用,肿瘤的照片图如图18所示,肿瘤的大体标准图如图19所示。从图18、图19可见,28天后对照组肿瘤体积最大,相同浓度下F/A-PLGA@DOX/SPIO组的肿瘤体积明显小于单独DOX组。通过游标卡尺测量并计算出相对肿瘤体积,结果如图20所示,从图20可见,高浓度F/A-PLGA@DOX/SPIO的肿瘤体积最少,并且低浓度F/A-PLGA@DOX/SPIO组的肿瘤体积也小于高浓度DOX组。图21、图22所示的相对肿瘤增殖率及肿瘤生长抑制率也符合上述结果,高浓度和低浓度组F/A-PLGA@DOX/SPIO抗肿瘤活性均高于高浓度DOX组。并且通过检测裸鼠体重发现(图23),DOX处理组给药6天内体重与对照组相比开始出现明显下降,到28天体重大约下降到17g,表明DOX对裸鼠的正常生长产生明显的影响。而F/A-PLGA@DOX/SPIO处理组的裸鼠在接受给药28天体重与对照组无明显差别,表明F/A-PLGA@DOX/SPIO对裸鼠的正常生长没有产生明显的影响,其毒副作用较小。通过核磁成像技术检测裸鼠肿瘤体积(图24),从图24可见,给药28天后F/A-PLGA@DOX/SPIO的肿瘤体积最小,远远小于对照组。
为了综合评价含聚合物纳米粒子对机体所产生的毒性,我们比较了药物注射28天后裸鼠心、肝、脾、肺、肾和肿瘤的组织切片变化和血液生化指标。将组织切片脱蜡至水后,在显微镜下观察组织状态,拍照。本发明实施例1所得聚合物纳米粒子治疗接种人肺癌细胞的荷瘤裸鼠心、肝、脾、肺、肾和肿瘤的H&E染色图如图25所示,我们发现DOX组的肺泡腔出血和肾小球萎缩的变化,而F/A-PLGA@DOX/SPIO组的脏器则未出现损伤或炎症反应。
28天治疗结束后,对裸鼠取血。完成取血后,血液立即使用3000rpm离心10min,收集血清,并置于碎冰中。从图26裸鼠血液指标图(同时设置未经人肺癌细胞A549接种且未给药的健康裸鼠为健康组)可见,DOX给药组明显地上调血脂相关指标总胆固醇(CHOL)、低密度脂蛋白胆固醇(LDL-C)、甘油三酯(TG),肝功能指标谷草转氨酶(AST),肾功能指标尿素氮(BUN)、肌酐(CREA)、尿酸(UA),心功能指标乳酸脱氢酶(LDH)、肌酸激酶(CK)。有可能是DOX的给药造成了裸鼠肾功能,肝功能和心功能的损伤。F/A-PLGA@DOX/SPIO组治疗后,肾功能相关的尿素氮(BUN)、肌酐(CREA)、尿酸(UA)渐趋正常水平。血液生化指标结果分析表明,F/A-PLGA@DOX/SPIO治疗后,有效降低肿瘤对小鼠血液生化指标的影响。其中,相对于DOX,靶向基团的引入使F/A-PLGA@DOX/SPIO能选择性地聚集在肿瘤组织,有效降低了DOX的肾毒性,明显下调了尿酸(UA)、尿素氮(BUN)和肌酐(CRE)。另外,F/A-PLGA@DOX/SPIO避免总胆固醇(CHOL)、低密度脂蛋白胆固醇(LDL-C)、甘油三酯(TG)的上升。
通过免疫组织化学法检测结果(图27),H&E染色可以发现,治疗组出现明显的坏死区,其中F/A-PLGA@DOX/SPIO 2mg/kg组最为明显。CD31又称为血小板-内皮细胞粘附分子,一般表达于血管内皮细胞,主要用于证明内皮细胞组织的存在,用于评估肿瘤血管生成情况,这可能意味着一个快速增长的肿瘤的程度,F/A-PLGA@DOX/SPIO组中,CD31阳性染色率明显低于其他各组。Ki-67也叫细胞增殖指数,是一种在细胞增殖G1,S,G2和M期中均出现的核抗原,由于其半衰期短,可以准确反映细胞的增殖活性,已广泛应用于多种肿瘤增殖活性的测定来判断肿瘤的恶性程度。F/A-PLGA@DOX/SPIO2mg/kg组中Ki67阳性染色率显著低于其他各组。VEGF又称血管内皮生长因子,是由不同肿瘤细胞分泌,可以促进血管内皮细胞增殖和新生血管形成、增加血管通透性、促进淋巴内皮细胞生长及增加组织因子的生成,是诱导肿瘤血管形成的作用最强、特异性最高的血管生成因子。F/A-PLGA@DOX/SPIO组中VEGF阳性染色显著高于其他各组。本结果进一步证明了F/A-PLGA@DOX/SPIO能够有抑制肿瘤细胞的生长,是一种高效低毒的纳米系统。综上所述,F/A-PLGA@DOX/SPIO通过靶向分子修饰后,能在荷瘤裸鼠模型中有效地被肿瘤组织吸收,从而提高其抗肿瘤活性和降低药物对正常组织的毒副作用。
上述实施例为本发明较佳的实施方式,但本发明的实施方式并不受上述实施例的限制,其他的任何未背离本发明的精神实质与原理下所作的改变、修饰、替代、组合、简化,均应为等效的置换方式,都包含在本发明的保护范围之内。

Claims (10)

1.一种磁共振成像纳米载体,其特征在于该载体为聚合物纳米粒子,所述的聚合物为PLGA-CS,其表面具有活性基团。
2.根据权利要求1所述的磁共振成像纳米载体,其特征在于:
所述的聚合物纳米粒子的尺寸为100~300纳米;
所述的活性基团为氨基、羟基或羧基。
3.根据权利要求1或2所述的磁共振成像纳米载体在制备抗肿瘤载药系统中的应用。
4.根据权利要求3所述的磁共振成像纳米载体在制备抗肿瘤载药系统中的应用,其特征在于所述的肿瘤包括肝癌、肺癌、恶性黑色瘤、乳腺癌、结肠癌、鼻咽癌、膀胱癌、宫颈癌、胃癌、食管癌、前列腺癌和结肠癌。
5.一种磁共振成像纳米载药系统,其特征在于载体为权利要求1或2所述的磁共振成像纳米载体。
6.根据权利要求5所述的磁共振成像纳米载药系统,其特征在于所述的载药系统主要由以下方法制备得到:
(1)将PLGA、核磁成像造影剂、抗肿瘤药物加入丙酮溶液中,得到PLGA丙酮溶液;
(2)将步骤(1)中得到的PLGA丙酮溶液逐滴加入吐温水溶液中,搅拌,得到PLGA水溶液;
(3)向步骤(2)中的得到的PLGA水溶液中加入N-羟基琥珀酰亚胺和1-(3-二甲氨基丙基)-3-乙基碳二亚胺盐酸盐活化PLGA上的羧基,再加入CS水溶液,搅拌,得到PLGA-CS溶液;
(4)向步骤(3)中得到的PLGA-CS溶液中加入靶向分子,搅拌,得到PLGA-CS聚合物纳米粒子水溶液,即形成磁共振成像纳米载药系统。
7.根据权利要求6所述的磁共振成像纳米载药系统,其特征在于:
步骤(1)中所述的抗肿瘤药物为柔红霉素、阿霉素、去甲氧柔红霉素、表阿霉素、紫杉醇、香菇多糖、长春花碱、长春新碱、三苯氧胺、福美司坦、阿那曲唑、氟他胺、5-氟尿嘧啶、甲氨蝶呤、顺铂、卡铂、奥沙利铂、卡莫司汀、托瑞米芬、替加氟、姜黄素、去甲氧基姜黄素、双曲甲氧基姜黄素和塞替派中的至少一种;
步骤(1)中所述的核磁成像造影剂为超小超顺磁性氧化铁纳米粒子或钆喷酸葡胺注射液;
步骤(1)中所得到的PLGA丙酮溶液中,PLGA的浓度为1~10mg/mL、核磁成像造影剂的浓度为1~20mg/mL、抗肿瘤药物的浓度为10~500μM。
8.根据权利要求6所述的磁共振成像纳米载药系统,其特征在于:
步骤(2)中所述的逐滴是指滴入速度为每滴间隔为1~10秒;
步骤(2)中所述的吐温水溶液是指浓度为1~5mg/mL的吐温水溶液;
步骤(2)中所述的搅拌的条件为200~800r/min搅拌过夜;
步骤(2)中所述的PLGA水溶液中抗肿瘤药物的浓度为4~200μM。
9.根据权利要求6所述的磁共振成像纳米载药系统,其特征在于:
步骤(3)中所加入的N-羟基琥珀酰亚胺与PLGA水溶液中PLGA的摩尔比为1~3:1;步骤(3)中所加入的1-(3-二甲氨基丙基)-3-乙基碳二亚胺盐酸盐与PLGA水溶液中PLGA的摩尔比为1~3:1;
步骤(3)中所述的活化PLGA上的羧基是指活化时间为2~12h;
步骤(3)中所述的CS水溶液的浓度为0.6~1mg/mL;所述的CS的分子量为5000~20000;所加入的CS水溶液的用量满足步骤(3)中PLGA水溶液中PLGA与CS水溶液中CS的摩尔比为1:1~10;
步骤(3)中所述的搅拌是指搅拌速度为200~800r/min,搅拌时间为8~24h。
10.根据权利要求6所述的磁共振成像纳米载药系统,其特征在于:
步骤(4)中所述的靶向分子为环形RGD多肽、叶酸、整合素、转铁蛋白、可活化细胞穿膜肽、MUC-1附膜蛋白、半乳糖胺、新生血管靶向肽、粒细胞巨噬细胞刺激因子中的两种;
步骤(4)中所述的靶向分子的添加量满足靶向分子与步骤(3)中加入的CS水溶液中CS的质量比为1:10;
步骤(4)中所述的搅拌是指在200~800r/min下搅拌过夜;
步骤(4)后还包括一个纯化步骤,所述的纯化步骤包括离心、重悬。
CN201810117789.7A 2018-02-06 2018-02-06 一种磁共振成像纳米载体、纳米载药系统及其制备方法 Active CN108404142B (zh)

Priority Applications (1)

Application Number Priority Date Filing Date Title
CN201810117789.7A CN108404142B (zh) 2018-02-06 2018-02-06 一种磁共振成像纳米载体、纳米载药系统及其制备方法

Applications Claiming Priority (1)

Application Number Priority Date Filing Date Title
CN201810117789.7A CN108404142B (zh) 2018-02-06 2018-02-06 一种磁共振成像纳米载体、纳米载药系统及其制备方法

Publications (2)

Publication Number Publication Date
CN108404142A true CN108404142A (zh) 2018-08-17
CN108404142B CN108404142B (zh) 2020-01-17

Family

ID=63127635

Family Applications (1)

Application Number Title Priority Date Filing Date
CN201810117789.7A Active CN108404142B (zh) 2018-02-06 2018-02-06 一种磁共振成像纳米载体、纳米载药系统及其制备方法

Country Status (1)

Country Link
CN (1) CN108404142B (zh)

Cited By (8)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN109432048A (zh) * 2018-11-22 2019-03-08 东华大学 一种血小板膜包裹载药多孔纳米颗粒及其制备方法
CN109536448A (zh) * 2018-12-10 2019-03-29 济宁医学院 一种多功能负载维甲酸的钆参杂四氧化三铁复合纳米粒子
CN111298139A (zh) * 2019-12-04 2020-06-19 暨南大学 基于核磁成像引导的克服肿瘤乏氧引起耐药性的靶向纳米药物及其制备方法和应用
CN111358954A (zh) * 2020-03-24 2020-07-03 中国人民解放军总医院 一种具备靶向调节巨噬细胞极化功能的组合物及其制备方法和用途
CN112791195A (zh) * 2021-02-24 2021-05-14 暨南大学 一种复合纳米载药体系的合成方法
CN112791184A (zh) * 2019-11-14 2021-05-14 美国发现集团有限公司 一种抗肿瘤纳米机器人及其制备方法
CN113975403A (zh) * 2021-11-03 2022-01-28 江苏护理职业学院 转铁蛋白与抗体结合超顺磁性氧化铁纳米粒及其制备方法和应用
CN114404429A (zh) * 2021-11-30 2022-04-29 重庆医科大学附属第二医院 一种纳米银修饰的单宁酸-铁网络的载药纳米复合物及其制备方法和逆转肿瘤耐药应用

Non-Patent Citations (2)

* Cited by examiner, † Cited by third party
Title
JAIME IBARRA等: ""Synthesis and characterization of magnetite/PLGA/chitosan nanoparticles"", 《MATER. RES. EXPRESS》 *
YICHAO WANG等: ""Chitosan-Modified PLGA Nanoparticles with Versatile Surface for Improved Drug Delivery"", 《AAPS PHARMSCITECH》 *

Cited By (14)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN109432048B (zh) * 2018-11-22 2021-03-19 东华大学 一种血小板膜包裹载药多孔纳米颗粒及其制备方法
CN109432048A (zh) * 2018-11-22 2019-03-08 东华大学 一种血小板膜包裹载药多孔纳米颗粒及其制备方法
CN109536448A (zh) * 2018-12-10 2019-03-29 济宁医学院 一种多功能负载维甲酸的钆参杂四氧化三铁复合纳米粒子
CN109536448B (zh) * 2018-12-10 2020-01-10 济宁医学院 一种多功能负载维甲酸的钆参杂四氧化三铁复合纳米粒子
CN112791184A (zh) * 2019-11-14 2021-05-14 美国发现集团有限公司 一种抗肿瘤纳米机器人及其制备方法
CN111298139B (zh) * 2019-12-04 2022-07-12 暨南大学 基于核磁成像引导的克服肿瘤乏氧引起耐药性的靶向纳米药物及其制备方法和应用
CN111298139A (zh) * 2019-12-04 2020-06-19 暨南大学 基于核磁成像引导的克服肿瘤乏氧引起耐药性的靶向纳米药物及其制备方法和应用
CN111358954A (zh) * 2020-03-24 2020-07-03 中国人民解放军总医院 一种具备靶向调节巨噬细胞极化功能的组合物及其制备方法和用途
CN112791195A (zh) * 2021-02-24 2021-05-14 暨南大学 一种复合纳米载药体系的合成方法
CN112791195B (zh) * 2021-02-24 2021-09-21 暨南大学 一种复合纳米载药体系的合成方法
CN113975403A (zh) * 2021-11-03 2022-01-28 江苏护理职业学院 转铁蛋白与抗体结合超顺磁性氧化铁纳米粒及其制备方法和应用
CN113975403B (zh) * 2021-11-03 2023-11-17 江苏护理职业学院 转铁蛋白与抗体结合超顺磁性氧化铁纳米粒及其制备方法和应用
CN114404429A (zh) * 2021-11-30 2022-04-29 重庆医科大学附属第二医院 一种纳米银修饰的单宁酸-铁网络的载药纳米复合物及其制备方法和逆转肿瘤耐药应用
CN114404429B (zh) * 2021-11-30 2023-06-30 重庆医科大学附属第二医院 一种纳米银修饰的单宁酸-铁网络的载药纳米复合物及其制备方法和逆转肿瘤耐药应用

Also Published As

Publication number Publication date
CN108404142B (zh) 2020-01-17

Similar Documents

Publication Publication Date Title
CN108404142A (zh) 一种磁共振成像纳米载体、纳米载药系统及其制备方法
CN107715121B (zh) 一种磁共振成像纳米药物载体、纳米载药系统及其制备方法
Li et al. Magnetic nanoparticles for cancer theranostics: Advances and prospects
Nelson et al. Use of superparamagnetic iron oxide nanoparticles (SPIONs) via multiple imaging modalities and modifications to reduce cytotoxicity: An educational review
Béalle et al. Ultra magnetic liposomes for MR imaging, targeting, and hyperthermia
Cao et al. Gadolinium-based nanoscale MRI contrast agents for tumor imaging
Luo et al. RGD-functionalized ultrasmall iron oxide nanoparticles for targeted T 1-weighted MR imaging of gliomas
Xie et al. Iron oxide nanoparticle platform for biomedical applications
Wahajuddin et al. Superparamagnetic iron oxide nanoparticles: magnetic nanoplatforms as drug carriers
Sun et al. PEG-mediated synthesis of highly dispersive multifunctional superparamagnetic nanoparticles: their physicochemical properties and function in vivo
Wang et al. In vivo dual-targeted chemotherapy of drug resistant cancer by rationally designed nanocarrier
Crayton et al. pH-titratable superparamagnetic iron oxide for improved nanoparticle accumulation in acidic tumor microenvironments
Du et al. Multi-functional liposomes showing radiofrequency-triggered release and magnetic resonance imaging for tumor multi-mechanism therapy
Hu et al. Facile synthesis of RGD peptide-modified iron oxide nanoparticles with ultrahigh relaxivity for targeted MR imaging of tumors
Zadeh et al. Smart pH-responsive magnetic graphene quantum dots nanocarriers for anticancer drug delivery of curcumin
Li et al. Aqueous-phase synthesis of iron oxide nanoparticles and composites for cancer diagnosis and therapy
Voulgari et al. Synthesis, characterization and in vivo evaluation of a magnetic cisplatin delivery nanosystem based on PMAA-graft-PEG copolymers
Hao et al. Multifunctional nanosheets based on folic acid modified manganese oxide for tumor-targeting theranostic application
Jing et al. Multifunctional nanoflowers for simultaneous multimodal imaging and high-sensitivity chemo-photothermal treatment
Shao et al. A smart multifunctional nanoparticle for enhanced near-infrared image-guided photothermal therapy against gastric cancer
Wang et al. Fe3O4@ Astragalus Polysaccharide Core–Shell Nanoparticles for Iron Deficiency Anemia Therapy and Magnetic Resonance Imaging in Vivo
Hao et al. Doxorubicin and Fe 3 O 4 loaded albumin nanoparticles with folic acid modified dextran surface for tumor diagnosis and therapy
Chen et al. Precise Study on Size‐Dependent Properties of Magnetic Iron Oxide Nanoparticles for In Vivo Magnetic Resonance Imaging
Chen et al. Improving sensitivity of magnetic resonance imaging by using a dual-targeted magnetic iron oxide nanoprobe
Patra et al. MRI‐visual order–disorder micellar nanostructures for smart cancer theranostics

Legal Events

Date Code Title Description
PB01 Publication
PB01 Publication
SE01 Entry into force of request for substantive examination
SE01 Entry into force of request for substantive examination
GR01 Patent grant
GR01 Patent grant