WO2010100108A1 - Procédé de fabrication d'un complexe de nanoparticules micellaires réticulés et produits de celui-ci - Google Patents

Procédé de fabrication d'un complexe de nanoparticules micellaires réticulés et produits de celui-ci Download PDF

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WO2010100108A1
WO2010100108A1 PCT/EP2010/052540 EP2010052540W WO2010100108A1 WO 2010100108 A1 WO2010100108 A1 WO 2010100108A1 EP 2010052540 W EP2010052540 W EP 2010052540W WO 2010100108 A1 WO2010100108 A1 WO 2010100108A1
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nanoparticle
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complex
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Elmar PÖSELT
Steffen Fischer
Stephan FÖRSTER
Horst Weller
Christian Schmidtke
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Centrum Für Angewandte Nanotechnologie (Can) Gmbh
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    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K49/00—Preparations for testing in vivo
    • A61K49/06—Nuclear magnetic resonance [NMR] contrast preparations; Magnetic resonance imaging [MRI] contrast preparations
    • A61K49/18—Nuclear magnetic resonance [NMR] contrast preparations; Magnetic resonance imaging [MRI] contrast preparations characterised by a special physical form, e.g. emulsions, microcapsules, liposomes
    • A61K49/1818—Nuclear 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
    • A61K49/1821—Nuclear 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
    • A61K49/1824—Nuclear 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
    • A61K49/1827—Nuclear 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
    • A61K49/1875—Nuclear 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 coated or functionalised with an antibody
    • A—HUMAN NECESSITIES
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    • A61K49/001—Preparation for luminescence or biological staining
    • A61K49/0013—Luminescence
    • A61K49/0017—Fluorescence in vivo
    • A61K49/0019—Fluorescence in vivo characterised by the fluorescent group, e.g. oligomeric, polymeric or dendritic molecules
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K49/00—Preparations for testing in vivo
    • A61K49/001—Preparation for luminescence or biological staining
    • A61K49/0063—Preparation for luminescence or biological staining characterised by a special physical or galenical form, e.g. emulsions, microspheres
    • A61K49/0065—Preparation for luminescence or biological staining characterised by a special physical or galenical form, e.g. emulsions, microspheres the luminescent/fluorescent agent having itself a special physical form, e.g. gold nanoparticle
    • A61K49/0067—Preparation for luminescence or biological staining characterised by a special physical or galenical form, e.g. emulsions, microspheres the luminescent/fluorescent agent having itself a special physical form, e.g. gold nanoparticle quantum dots, fluorescent nanocrystals
    • A—HUMAN NECESSITIES
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    • A61K49/00—Preparations for testing in vivo
    • A61K49/001—Preparation for luminescence or biological staining
    • A61K49/0063—Preparation for luminescence or biological staining characterised by a special physical or galenical form, e.g. emulsions, microspheres
    • A61K49/0069—Preparation for luminescence or biological staining characterised by a special physical or galenical form, e.g. emulsions, microspheres the agent being in a particular physical galenical form
    • A61K49/0076—Preparation for luminescence or biological staining characterised by a special physical or galenical form, e.g. emulsions, microspheres the agent being in a particular physical galenical form dispersion, suspension, e.g. particles in a liquid, colloid, emulsion
    • A61K49/0082—Preparation for luminescence or biological staining characterised by a special physical or galenical form, e.g. emulsions, microspheres the agent being in a particular physical galenical form dispersion, suspension, e.g. particles in a liquid, colloid, emulsion micelle, e.g. phospholipidic micelle and polymeric micelle
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
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    • A61K49/00—Preparations for testing in vivo
    • A61K49/001—Preparation for luminescence or biological staining
    • A61K49/0063—Preparation for luminescence or biological staining characterised by a special physical or galenical form, e.g. emulsions, microspheres
    • A61K49/0069—Preparation for luminescence or biological staining characterised by a special physical or galenical form, e.g. emulsions, microspheres the agent being in a particular physical galenical form
    • A61K49/0089—Particulate, powder, adsorbate, bead, sphere
    • A61K49/0091—Microparticle, microcapsule, microbubble, microsphere, microbead, i.e. having a size or diameter higher or equal to 1 micrometer
    • A61K49/0093—Nanoparticle, nanocapsule, nanobubble, nanosphere, nanobead, i.e. having a size or diameter smaller than 1 micrometer, e.g. polymeric nanoparticle
    • A—HUMAN NECESSITIES
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    • A61K49/06—Nuclear magnetic resonance [NMR] contrast preparations; Magnetic resonance imaging [MRI] contrast preparations
    • A61K49/18—Nuclear magnetic resonance [NMR] contrast preparations; Magnetic resonance imaging [MRI] contrast preparations characterised by a special physical form, e.g. emulsions, microcapsules, liposomes
    • A61K49/1806—Suspensions, emulsions, colloids, dispersions
    • A—HUMAN NECESSITIES
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    • A61K49/06—Nuclear magnetic resonance [NMR] contrast preparations; Magnetic resonance imaging [MRI] contrast preparations
    • A61K49/18—Nuclear magnetic resonance [NMR] contrast preparations; Magnetic resonance imaging [MRI] contrast preparations characterised by a special physical form, e.g. emulsions, microcapsules, liposomes
    • A61K49/1818—Nuclear 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
    • A61K49/1821—Nuclear 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
    • A61K49/1824—Nuclear 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
    • A61K49/1827—Nuclear 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
    • A61K49/1851—Nuclear 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
    • A61K49/1854—Nuclear 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 by reactions only involving carbon-to-carbon unsaturated bonds, e.g. poly(meth)acrylate, polyacrylamide, polyvinylpyrrolidone, polyvinylalcohol
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K49/00—Preparations for testing in vivo
    • A61K49/06—Nuclear magnetic resonance [NMR] contrast preparations; Magnetic resonance imaging [MRI] contrast preparations
    • A61K49/18—Nuclear magnetic resonance [NMR] contrast preparations; Magnetic resonance imaging [MRI] contrast preparations characterised by a special physical form, e.g. emulsions, microcapsules, liposomes
    • A61K49/1818—Nuclear 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
    • A61K49/1821—Nuclear 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
    • A61K49/1824—Nuclear 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
    • A61K49/1827—Nuclear 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
    • A61K49/1851—Nuclear 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
    • A61K49/1857—Nuclear 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
    • A61K49/186—Nuclear 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 the organic macromolecular compound being polyethyleneglycol [PEG]
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K49/00—Preparations for testing in vivo
    • A61K49/06—Nuclear magnetic resonance [NMR] contrast preparations; Magnetic resonance imaging [MRI] contrast preparations
    • A61K49/18—Nuclear magnetic resonance [NMR] contrast preparations; Magnetic resonance imaging [MRI] contrast preparations characterised by a special physical form, e.g. emulsions, microcapsules, liposomes
    • A61K49/1818—Nuclear 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
    • A61K49/1887—Agglomerates, clusters, i.e. more than one (super)(para)magnetic microparticle or nanoparticle are aggregated or entrapped in the same maxtrix
    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B82—NANOTECHNOLOGY
    • B82Y—SPECIFIC USES OR APPLICATIONS OF NANOSTRUCTURES; MEASUREMENT OR ANALYSIS OF NANOSTRUCTURES; MANUFACTURE OR TREATMENT OF NANOSTRUCTURES
    • B82Y30/00—Nanotechnology for materials or surface science, e.g. nanocomposites
    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B82—NANOTECHNOLOGY
    • B82Y—SPECIFIC USES OR APPLICATIONS OF NANOSTRUCTURES; MEASUREMENT OR ANALYSIS OF NANOSTRUCTURES; MANUFACTURE OR TREATMENT OF NANOSTRUCTURES
    • B82Y5/00—Nanobiotechnology or nanomedicine, e.g. protein engineering or drug delivery
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
    • G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
    • G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
    • G01N33/58—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving labelled substances
    • G01N33/585—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving labelled substances with a particulate label, e.g. coloured latex
    • G01N33/587—Nanoparticles

Definitions

  • the field of the present invention relates to a method for the manufacture of a cross-linked micellar nanoparticle complex which comprises a cross-linkage between a polymeric ligand and products thereof.
  • the cross-linked micellar nanoparticle complex has applications in biological systems.
  • US 2004142578 is owned by Ulrich Wiesner, Phong Du, Charles Black and Kathryn Guarayni (USA) and is titled, "Thin Film nanostructures".
  • the Wiesner et al patent application discloses the co-self- assembly of an organic (e.g., block copolymer) and an inorganic (e.g., sol-gel) component to create nano-metre features of silicon dioxide materials in thin films on silicon surfaces.
  • the '578 invention uses sol-gel chemistry to introduce an inorganic component, preferably 3-glycidoxy-propyltrimethoxysilane and aluminum- tri-sec-butoxide into a block copolymer, preferably poly (isoprene-block-ethylene oxide)-(PI-b-PEO), as a structure-directing agent.
  • the inorganic components preferentially migrate to the PEO block and swell the copolymer into different morphologies depending upon the amount of sol-gel precursors added.
  • Thin films with a thickness of below 100 nm are created by spin coating the hybrid solution onto a silicon wafer.
  • An inverse hexagonal morphology for example, is produced in which the polymer forms nano-pores within an inorganic matrix.
  • the monomers attached to the nanoparticles are polymerized to form a polymer layer on the individual nanoparticles within the discrete aqueous regions.
  • the polymerization comprises adding a cross-linker to the solution to cross-link the monomers attached to the individual nanoparticles.
  • the solution for coating individual nanoparticles may comprise a micro-emulsion comprising a continuous phase and a discrete aqueous region defined by reverse micelles; hydrophobic nanoparticles dispersed in the micro-emulsion; amphiphilic polymerisable monomers attachable to the hydrophobic nanoparticles; and a cross-linker for polymerizing the monomers.
  • WO2007/018647 by Yang et al. is titled "Multifunctional nanostructures, methods of synthesising thereof and methods of use thereof.
  • the WO2007/018647 document describes nanostructure including a nano- species, a hydrophobic protection structure including at least one compound selected from a capping ligand, an amphiphilic copolymer, and combinations thereof, wherein the hydro- phobic protection structure encapsulates the nano-species, and at least one histidine-tagged peptide or protein conjugated to the hydrophobic protection structure, wherein the at least one histidine-tagged peptide or protein has at least one binding site.
  • WO2009/061456 document describes a biomimetic contrast agent comprising an amine- functionalized iron (II) oxide/iron(lll) oxide nanoparticle core a targeting ligand attached to -A- the nanoparticle core via a linker and an inert outer layer of a hydrophilic polymer conjugated to the targeting ligand and imaging methods using the biomimetic contrast agents.
  • US patent application publication No. US2004/115433 by Elaissari et al. is titled "Composite particles, derived conjugates, preparation method thereof and applications of same".
  • the US2004/115433 document describes composite particles comprising a hydrophobic polymer core and inorganic nanoparticles.
  • the hydrophobic polymer forms a polymer matrix inside which the inorganic nanoparticles are stabilized and distributed in a relatively homogenous manner.
  • the inorganic nanoparticles are at least partially surrounded by an amphophilic copolymer comprising a hydrophobic part and a hydrophilic part, said hydrophobic part being at least partially immobilized in the polymer matrix.
  • WO2005/102396 by Nie and Gao is titled "Multimodality nanostructures, methods of fabricating thereof, and methods of use thereof.
  • the WO2005/102396 document describes a multimodality nanostructure, comprising nano-species having a first detectable functionality; a second detectable functionality, wherein the second molecule is attached to the nano-species, and wherein the first detectable functionality and the second detectable functionality are different.
  • WO2008/054523 by Yang et al. is titled "Multifunctional nanostructures, methods of synthesising thereof and methods of use thereof.
  • the WO2008/054523 document describes a nanostructure that includes a magnetic iron oxide nanoparticle, a hydrophobic protection structure including at least an am- phiphilic copolymer, wherein the hydrophobic protection structure encapsulates the mag- netic iron oxide nanoparticle and at least one amino-terminal fragment (ATF) peptide or epidermal growth factor receptor (EGFR) antibody conjugated to the amphiphilic copolymer.
  • ATF amino-terminal fragment
  • EGFR epidermal growth factor receptor
  • Nanoparti- cles have applications in a wide range of technical fields. Nanoparticles have applications amongst others in the technical field of sensing, fluorescent labelling, optical imaging, magnetic resonance imaging, cell separation, and for the treatment of diseases in biomedical applications. Nanoparticles also have applications in the technical field of chemical catalysis, stabilisers for dyes and uses in solar cells.
  • the nanoparticle When the nanoparticle is used in biomedical applications, the nanoparticle must exhibit a degree of water solubility and rigidity towards an aqueous environment which is found in the biomedical application. Poor solubility of potentially useful nanoparticles for biomedical applications is a problem when the nanoparticle has a hydrophobic nature.
  • the nanoparticle may be capped with an organic ligand layer to manufacture a nanoparticle complex which has a degree of water solubility (see for example P. Alivisa- tos, Nature Biotechnology 2004, 22, 47 and X. Michalet, F. F. Pinaud, L. A. Bentolila, J. M.
  • the hydrophilic organic ligand used to manufacture the water-soluble nanoparticle complex usually posses nucleophilic groups such as thiols, amines, and phosphines to coordinate to the nanoparticle surface. Further functional groups at the other end of the hydrophilic organic ligand, which are to the outside of the nanoparticle complex directed, are important when the nanoparticle is used in biomedical applications because the functional groups render the nanoparticle more selective to certain biological targets when the nanoparticle is used in the biomedical application.
  • the hydrophilic organic ligand affects the physical properties of the nanoparticle.
  • the ligand exchange of the organic ligand layer with the hydrophilic organic ligand can lead to a drop in quantum yield of the quantum dot nanoparticle.
  • the drop in quantum yield of the quantum dot nanoparticle can result from a reduced passivation layer on the nanoparticle (see for example H. Fan, E. W. Leve, C. Scullin, J. Gabaldon, D. Tallant, S. Bunge, T. Boyle, M. C.Wilson, C. J. Brinker, Nano Letters 2005, 5, 645).
  • a different approach to achieve a degree of water solubility of the nanoparticle is to provide a micellar coating to the nanoparticle to manufacture a capped micellar nanopar- tide complex.
  • the micellar coating of the nanoparticle is known to conserve the hydrophobic ligand shell of a quantum dot nanoparticle (see for example H. Fan, E. W. Leve, C. Scullin, J. Gabaldon, D. Tallant, S. Bunge, T. Boyle, M. C.Wilson, C. J. Brinker, Nano Lett. Nano Letters 2005, 5, 645) thus reducing any loss in quantum yield of the quantum dot nanoparticle.
  • Nanoparticles have been coated with a block copolymer ligand to manufacture the capped micellar nanoparticle.
  • the use of the block copolymer ligand provides a means to manufacture the capped micellar nanoparticle complex which is sufficiently stable for in vivo applications and which therefore have potential for use in biomedical applications (see for example Z. Gao, A. N. Lukyanov, A. Singhal, V. P. Torchilin, Nano Lett. 2002, 2, 979).
  • a method for the manufacture of the block copolymer ligand is to use a selective solvent (see for example M. Svensson, P. Alexandridis, P. Linse, Macromolecules 1999, 32, 637).
  • the block copolymer ligand is usually amphiphilic.
  • the selective solvent is selective for one of the two copolymer blocks during the manufacture of the block copolymer ligand.
  • aggregation of the non- soluble block leads to the formation of the micelle with predictable sizes and morphologies.
  • Dual interaction ligands provide an option for the manufacture of capped micellar nanoparticle for use in biomedical applications.
  • the dual interaction ligands can utilise hydrophobic van der Waals interactions and utilise coordinate bonding for the manufacture of the micellar nanoparticle complex to overcome the problems mentioned above.
  • the dual interaction ligand should provide the possibility for coupling of the dual interaction ligand to, for example, antibodies and provide the possibility that the capped micellar nanoparticle should be able to maintain a high flow rate by avoiding ag- gregations for example in the blood stream for in vivo biomedical applications.
  • Dual interaction ligands are known, wherein a covalent bond is used to cross-link polymer blocks around the nanoparticle (see for example V. Buetuen, X. S. Wang, M. V. d. P. Banez, K. L. Robinson, N. C. Billingham, S. P. Armes, Z. Tuzar, Macromolecules 2000, 33, 1 and Q. Zhang, E. E. Remsen, K. L. Wooley, J. Am. Chem. Soc. 2000, 122, 3642).
  • the ligand shell that surrounds the nanoparticle should be cross linked by a cross -linkage.
  • the cross-linkage should not de- crease the flexibility of the hydrophilic ligand (-segment).
  • the number of trap states can be decreased by an inorganic passivation layer, which has larger band gaps like ZnS (see Figure 1).
  • CdS is been used as interlayer to reduce the strain regarding the lattice mismatch between the CdSe and ZnS.
  • the trapped electrons can support the non-radiative recombining of other electron-hole pairs (excitons) and elongate the dark state of the nanoparticle. Therefore when considering an inhomoge- neous inorganic passivation layer a dipolar or electrostatic interaction between a ligand, a solvent and the quantum dot nanoparticle becomes relevant.
  • the present invention discloses a method for the manufacture of a cross-linked micellar nanoparticle complex, wherein a cross -linkable amphiphilic block copolymer is cross-linked to a form a stable micellar nanoparticle.
  • the cross-linked micellar nanoparti- cle complex maintains water solubility of the nanoparticle with a hydrophilic outer micellar structure while a hydrophobic, cross-linked inner structure assures the preservation of the ligand shell in an aqueous medium.
  • the cross-linked micellar nanoparticle complex conserves the electrical and optical properties of the nanoparticle while preserving the flexibility of the outermost polymer of the micellar cross-linked nanoparticle complex.
  • the cross -linkable amphiphilic block copolymer allows the modification of terminal groups of the amphiphilic block copolymer.
  • the functionalisation of the terminal groups of the amphiphilic block copolymer facilitates the use of the cross-linked micellar nanoparticle complex in biomedical applications.
  • the cross-linked micellar nanoparticle complex comprises the capped micellar nanoparticle, which has intermolecular cross linkages from the cross-linkable polymer ligand which is attached to the nanoparticle, to the cross -linkable amphiphilic block copolymer.
  • the cross-linked micellar nanoparticle complex has at least one nanoparticle present in the cross-linked micellar nanoparticle complex.
  • the cross-linked micellar nanoparticle complex comprises terminal functional groups on the cross-linkable amphiphilic block copolymer.
  • micellar nanoparticle complex is soluble in aqueous media and can be used in biomedical applications. Description of Figures
  • Figure 1 illustrates trap states located on the surface of a core/shell/shell semiconducting nanoparticle with an inhomogeneous shell. On the left are the lattice mismatches and the band gaps shown of the CdSe, CdS, and ZnS.
  • Figure 2 shows a 1 H-NMR spectra of a cross linkable polymer ligand (PI-N3) and a cross -linkable amphiphilic block copolymer (PI O1 -ID-PEO 2I2 -OH) according to an aspect of the present invention.
  • PI-N3 cross linkable polymer ligand
  • PI O1 -ID-PEO 2I2 -OH cross -linkable amphiphilic block copolymer
  • Figure 3 shows a schematic for the manufacture of a cross-linked nanoparticle complex according to the present invention.
  • Figure 4 shows dynamic light scattering measurements (left) of a CdSe/CdS/ZnS core/shell/shell capped micellar nanoparticle with a cross linkable polymer ligand PI 30 -N3 and a corresponding TEM image of the capped micellar nanoparticle (right).
  • FIG. 5 dynamic light scattering measurements (left) of a cross-linked nanoparti- cle complex 50 according to an example of the present invention and the corresponding TEM image of the capped micellar nanoparticle (right).
  • Figure 6 shows a schematic of a cross-linked nanoparticle complex 50, with more than one nanoparticle core (left) and with a single nanoparticle core (right).
  • Figure 7 shows a TEM image of capped micellar nanoparticle capped with the PINS and PI-PEO cross linkable polymer ligands.
  • the figure to the left shows magnification bar - 500 nm and the figure to the right shows magnification bar - 200 nm.
  • Figure 8 shows the results of dilution series measurements for measuring the emission of a capped micellar nanoparticle ( Figure 8A) and a cross-linked micellar nanoparticle complex (Figure 8B).
  • Figure 9 shows the fluorescence yield of a cross-linked micellar nanoparticle complex under various pH conditions and with various surfactants, based on the value in water.
  • Figure 10 shows the absorption, emission and dynamic light scattering results of a cross-linked micellar nanoparticle complex under various conditions of pH and with various surfactants over a period of 10 days.
  • Figure 11 shows the 1 H-NMR spectra of a cross -linkable amphiphilic block copolymer PI O1 -ID-PEO 212 -OH (above) in comparison to a cross -linkable amphiphilic block copolymer PI 61 -b-PEO 212 -COOH (below).
  • Figure 12 shows a FT-IR spectrum of a cross-linkable amphiphilic block copoly- mer PI O1 -ID-PEO 212 -OH in comparison to a cross-linkable amphiphilic block copolymer of PI 61 -b-PEO 212 -COOH.
  • Figure 13 shows a 1 H-NMR spectrum of a cross-linkable amphiphilic block copolymer PI 61 -b-PEO 212 -CHO.
  • Figure 14 shows a FT-IR spectrum of PI O1 -ID-PEO 212 -OH in comparison to PI 61 -ID- PEO 212 -ALKYNE.
  • Figure 15 shows the 1 H-NMR spectrum of a cross -linkable amphiphilic block copolymer PI 61 -b-PEO 212 - ALKYNE.
  • Figure 16 shows the 1 H-NMR spectrum of a cross -linkable amphiphilic block copolymer PI 61 -b-PEO 212 - ALKENE.
  • Figure 17 shows the 1 H-NMR spectrum of a cross -linkable amphiphilic block copolymer PI 61 -b-PEO 212 - AMINE.
  • Figure 18 shows a 1 H-NMR spectrum of a cross-linkable amphiphilic block copolymer PI 61 -b-PEO 212 -BORONIC ACID-ESTER.
  • Figure 19 shows a 1 H-NMR spectrum of a cross-linkable amphiphilic block co- polymer PI 61 -b-PEO 212 -EPOXIDE.
  • Figure 20 shows a 1 H-NMR spectrum of a cross-linkable amphiphilic block copolymer PI 61 -b-PEO 212 -galactopyranosid.
  • Figure 21 shows MRI images of a mouse exposed to a cross-linked micellar Fe 3 O 4 nanoparticle complex that present an anti-YKL-40 antibody on the surface and a cross- linked micellar quantum dot that present an anti-YKL-40 antibody on the surface too.
  • Figure 22 shows MRI images of a mouse exposed to the IgG2b-isotype-quantum dot and - ferric oxide conjugates.
  • Figure 23 shows confocal microscope images of a histological transaction of a tumour tissue, A: bonded antibody anti-YKL-40-quantum dot and B: tumour tissue of the isotype control.
  • Figure 24 shows the potential use of a cross-linked micellar nanoparticle complex for use as a drug delivery system.
  • Figure 25 shows a 1 H-NMR spectrum of a cross-linkable amphiphilic block co- polymer, PI 61 -b-PEO 212 -COOH coupled to Alexa Fluor® 594.
  • biomedical application includes, but is not limited to, diagnostics, therapeutics, in vivo, in vitro, living biological tissues, post mortem tissues.
  • the application of a nanoparticle 5 in an aqueous environment provides certain challenges due to the hydrophobic nature of the nanoparticle 5 and a loss in fluorescence quantum yield which is associated with the organic and inorganic passivation layer (as discussed above).
  • Polar solvents such as water, are known to stabilise electrons in trap states.
  • the trap states can lead to non-radiative processes. Due to a release of vibration energy the electron relaxes again to a trap ground state.
  • the surface of the pas- sivated nanoparticle 5 could become oxidised.
  • the oxidisation of the surface of the pas- sivated nanoparticle 10 leads to a loss of the inorganic passivation shell on the surface of the nanoparticle 5.
  • the inhomogeneous surface of the nanoparticle 5 affects the toxicity and the optical and the physical properties of the nanoparticle 5.
  • An intermolecular covalent cross-linkage 60 is formed between a cross -linkable polymer ligand 20 which is attached to the nanoparticle 5 to form a capped nanoparticle 30 and between a cross-linkable amphiphilic block copolymer 40.
  • the intermolecular covalent cross-linkage 60 is achieved in the inner segment of the micellar nanoparticle complex 50.
  • the intermolecular covalent cross- linkage of the inner Pi-blocks affords the flexibility of the outer PEO blocks.
  • the cross-linkable polymer ligand 20 which comprises a poly (isoprene) polymer with a primary terminal amine function was manufactured to bind to the nanoparticle 5 by the primary terminal amine function of the cross -linkable polymer ligand 20.
  • the cross-linkable polymer ligand 20 achieves a packing around the nanoparticle 5 to form the capped nanoparticle 30.
  • the cross-linkable amphiphilic block copolymer 40 was manufactured.
  • the cross- linkable amphiphilic block copolymer 40 comprises a hydrophobic poly (isoprene) (PI) block and a hydrophilic poly (ethylene oxide) (PEO) block.
  • PI poly (isoprene)
  • PEO poly (ethylene oxide)
  • the cross -linkable amphiphilic block copolymer 40 is PI O1 -ID-PEO 212 -OH and comprises a hydrophilic (PEO) block weight fraction of 70% of the total mass of the cross-linkable amphiphilic block copolymer 40.
  • Figure 2 shows the 1 H-NMR spectra of the cross -linkable amphiphilic block copolymer 40 Pl 6 i-b-PEO 212 -OH (top), PI O1 -OH (middle) and the cross linkable polymer ligand 20 PI 30 -N3 (bottom).
  • the cross-linked micellar nanoparticle complex 50 is manufactured (see Figure 3).
  • the critical micelle concentration is the concentration above which micelles are spontaneously formed.
  • the cross-linked micellar nanoparticle complex 50 has a hydrophobic packing around the core of the nanoparticle 5.
  • a cross-linkage initiator 70 is present.
  • the cross-linkage initiator 70 facilitates the cross-linkage between the cross -linkable polymer ligand 20 of the capped micellar nanoparticle 30 and the cross-linkable amphi- philic block copolymer 40.
  • Several CdSe/CdS/ZnS core/shell/shell passivated nanoparticles 10 and other nanoparticles like iron oxide, GdPO 4 , and gold were equipped with the cross -linkable polymer ligand 20 PI 30 -N3 to manufacture the capped nanoparticle 30 by a ligand exchange process.
  • a characteristic of a disposed TOP/TOPO organic passivation ligand layer of the nanoparticle 10 is a steady quantum yield of the nanoparticle 5 in the THF aside from a maximal drop in quantum yield of 10% shortly after dilution in the THF.
  • the cross- linkable polymer ligand 20 PI 30 -N3 ligand serves on one hand as an organic passivation ligand layer of the trap state and on the other hand for the conservation of the nanoparticle 5.
  • Dynamic light scattering (DLS) measurements of stock solutions of the capped nanoparticle 30 in chloroform shows (see Figure 4) a hydrodynamic diameter with a maximum in the intensity PSD distribution between 10 and 17 nm, primarily depending on the length of the PI block.
  • the PI block comprises of 50 monomer units.
  • the presence of the solvated non-cross-linked PEO block of the cross-linkable amphiphilic block copolymer 40 prevents the formation of a macro-gel and leads to the formation of the cross-linked micellar nanoparticle complex 50.
  • the initiation of the cross- linkage between the cross -linkable polymer ligand 20 of the capped micellar nanoparticle 30, with the cross-linkable amphiphilic block copolymer 40 can be provided by the use of a cross-linkage initiator 70, such as a radical initiator like AIBN or benzoyl peroxide or IRGACUR. The initiation can be achieved, depending on the initiator, thermally or by UV- light.
  • Electron Spin Resonance, IR and NMR measurements of the PI block of the cross -linkable polymer ligand 20 and the cross -linkable amphiphilic block copolymer 40 after UV-irradiation reveals the presence of unpaired electrons.
  • the presence of unpaired electrons signifies magnetic dipole/dipole interactions which shows a loss of double bounds, hydrogen formation, and therefore the presence of the intermolecular cross linkage
  • the intramolecular cross linkage 60 is conveying by the presence of isoprene.
  • the PI block of the cross -linkable polymer ligand 20 and the cross -linkable amphiphilic block copolymer 40 become closer during the formation of the intermolecular cross linkage 60 and consequently the manufacture of the cross-linked micellar nanoparticle complex 50.
  • the parameters include the properties of the cross-linkable amphiphilic block copolymer 40, the passivated nanoparticle 10 to cross- linkable polymer ligand 20 ratio and the amounts of any solvents added and the concentration of the THF solution of the cross-linked micellar nanoparticle complex 50 before injection of the mixture in water as well as the speed of injection.
  • micellar nanoparticle complex It is also known that the faster water surrounds the ligand-nanoparticle-THF solution the faster the manufacture of the micellar nanoparticle complex occurs. A faster rate of manufacture of the micellar nanoparticle complex results in a corresponding decrease in mean diameter of the cross -linkable micellar nanoparticle complex, which generates the cross-linked micellar nanoparticle complex 50 after cross linkage.
  • the cross-linked micellar nanoparticle complex 50 can comprise a cluster of the nanoparticles 5 in the cross-linked micellar nanopar- ticle complex 50.
  • the formation of the cluster of the nanoparticles 5 within the cross- linked micellar nanoparticle complex 50 depends on a ratio of the capped nanoparticle 30 to the cross-linkable amphiphilic block copolymer 40, during the injection in water.
  • Figure 6 (left) illustrates the cross-linked micellar nanoparticle complex 50 with the cluster of the nanoparticles 5 in the cross-linked micellar nanoparticle complex 50.
  • the cross-linked micellar nanoparticle complex 50 can comprise a cluster of nanoparticles 5 in the cross-linked micellar nanoparticle complex 50 (see Figure 6, left) or the cross-linked micellar nanoparticle complex 50 can comprise a single nanoparticle 5, in the cross-linked micellar nanoparticle complex 50 (see Figure 6, right).
  • the clustering of nanoparticles 5 in the cross-linked micellar nanoparticle complex 50 allows for the cross-linked micellar nanoparticle complex 50 to exhibit enhanced properties. It has been reported that clustering of super-paramagnetic magnetite nanoparticles 5 results in a saturated and higher magnetisation than that of individual super- paramagnetic magnetite nanoparticles 5. The saturated and higher magnetisation of the super-paramagnetic magnetite nanoparticles 5 arises due the interaction between the clustered super-paramagnetic magnetite nanoparticles 5.
  • the cross-linked micellar nanoparticle complex 50 that comprises clustered super-paramagnetic magnetite nanoparticles 5 can have applications in magnetic separation, drug delivery and cancer hyperthermia magnetite nanoparticles and is therefore a promising material for use in magnetic resonance imaging (MRI) contrast agents (see for example H. Gu, K. Xu, C. Xu, B. Xu, Chemical Comm. 2006, 941 and J.-H. Lee, Y.-w. Jun, S.-I. Yeon, J.-S. Shin, J. Cheon, Angewandte Chemie, International Edition 2006, 45, 8160).
  • MRI magnetic resonance imaging
  • the manufacture of the cross-linked micellar nanoparticle complex 50 that comprises different nanoparticles 5 in the cross-linked micellar nanoparticle complex 50 provides another route for enhanced properties of the cross-linked micellar nanoparticle complex 50.
  • the presence of magnetic nanoparticles 5 are known to quench CdSe/ZnS core/shell nanoparticles 5 by dynamic quenching (see for example K. Mandal Swapan, N. Lequeux, B. Rotenberg, M. Tramier, J. Fattaccioli, J. Bibette,B. Dubertret, Langmuir surfaces and colloids 2005, 21, 4175 and T. R. Sathe, A. Agrawal, S. Nie, Anal. Chem. 2006, 78, 5627).
  • micellar nanoparticle 30 comprising the CdSe/CdS/ZnS core/shell/shell nanoparticles 5 and the PINS cross -linkable polymer ligand 20. It has been shown that the length of the PI block of the cross -linkable polymer ligand 20 constitutes the interspace between the magnetic nanoparticles 5 and the CdSe/CdS/ZnS core/core/shell nanoparticles 5.
  • nanoparticles in addition to magnetic nanoparticles all kinds of different nanoparticles as well as different sized nanoparticles can be co-encapsulated in or on the micelle. Regarding the properties of for example gold, GdPO 4 , NaYF 4 and further nanoparticles this opens options for combining different physical properties of the nanoparticles.
  • quantum dot nanoparticles 5 make the quantum dot nanoparticles 5 suitable for use in multicolour imaging and in optical multiplexing applications.
  • the multiple colours and intensities can be combined to encode genes, proteins and small-molecule libraries.
  • the cross-linked micellar nanoparticle complex 50 showed a greater firmness.
  • the greater firmness is due to the intermolecular cross-linkage 60 between the PI blocks of the cross -linkable amphiphilic block copolymer 40 and the PI blocks of the cross -linkable polymer ligand 20.
  • the nanoparticle 5 of the cross-linked micellar nanoparticle complex 50 shows a steady quantum yield in water.
  • the steady quantum yield of the nanoparticle 5 of the cross-linked micellar nanoparticle complex 50 in water indicates that little of the cross -linkable polymer ligand 20 and 40 has been removed from a surface of the nanoparticle 5 and that a successful intermolecular cross-linkage 60 has been formed.
  • the intermo- lecular cross-linkage 60 was confirmed on the basis of stability tests on the cross-linked micellar nanoparticle complex 50.
  • the cross-linked micellar nanoparticle complex 50 was lyophilised and re-suspended either in the THF solution or in chloroform. In the THF solution or in chloroform the hydrophobic Pi-block is next to the PEO-block soluble and therefore any capped micellar nanoparticle complex 50 would be unstable. A marginal swelling of the particle diameter is detectable by DLS measurements, while no capped micellar nanoparticle complex 50 precipitated after four days.
  • micellar nanoparticle complex 50 To further determine the rigidity of the cross-linked micellar nanoparticle complex 50 various stability tests were conducted on the cross-linked micellar nanoparticle complex 50. The stability tests were conducted at various pH values of between 4 and 9 and by the addition of various surfactants. It is known that in the presence of surfactants, interactions occur with a micellar structure which often hinders the micellar structure.
  • the non-ionic surfactant pentaethylene glycol dodecyl ether is known to interact with a micellar structure that comprises a polybutadienepoly (ethylene oxide) (PB-PEO) copolymer, resulting in changes in the geometry of the micellar structure.
  • PB-PEO polybutadienepoly
  • Ionic surfactants such as sodium dodecyl sulphate (SDS) and dodecyltrimethylammonium bromide (DTAB) also interact with the micellar structure which hinders the formation of the micellar structure.
  • SDS sodium dodecyl sulphate
  • DTAB dodecyltrimethylammonium bromide
  • the interaction of the surfactants was tested on the cross-linked micellar nanoparticle complex 50 that comprises the capped micellar nanoparticle 30 with a CdSe/CdS/ZnS core/shell/shell nanoparticle 5 and the cross -linkable amphiphilic block copolymer 40 Pl ⁇ i- b-PEO 2 i 2 -OH.
  • the CdSe/CdS/ZnS core/shell/shell nanoparticle 5 had a diameter of ⁇ 3 nm, which exhibits a higher surface to volume ratio which makes the nanoparticle 5 more sensitive to environmental changes because trap states become more relevant.
  • the trap states can be populated by electrons and quench the quantum yield of the quantum dot nanoparticle 5.
  • 1.7 • 10 "9 mol of the nanoparticles 5 (nanoparticle: Pl ⁇ i-b- PEO 2I2 -OH ratio 1:100) was dissolved in 3 mL of an aqueous solution containing the sur- factant (DTAB, NaCl, EDTA, and SDS) or different pH values (4-9).
  • the capped micellar nanoparticle 30 comprising the nanoparticle 5 provides a local chemical environment similar to that of the initial organic solvent.
  • the inert PEO block of the cross-linkable polymer ligand 20 ensures the steadiness of the capped micellar nanoparticle 30 against biological degradation. Not only the tightness of the cross -linkable polymer ligand 20, but also the feasibility to use low con- centrations of the CdSe nanoparticle 5, has to be reflected in the reduced cytotoxicity.
  • the PEO block has the ability to prevent protein adsorption, cell adhesion and consequently phagocytosis, non-initiation of an immune response, thus, increasing the in vivo circulation lifetime (serum lifetime).
  • PEO-conjugation of enzymes and other biomolecules with the preservation of the enzymes biological activity is known (see for Example J. A. Neff, K. D. Caldwell, P. A. Tresco, J. Biomed. Mater. Res. 1998, 40, 511).
  • the cross-linkable amphiphilic block copolymer 40 PI O1 -ID-PEO 2I2 -OH possesses just one terminal functional group, the -OH group.
  • the -OH terminal functional group provides a means for the functionalisation of the cross -linkable amphiphilic block copolymer 40 Pl 6 i-b-PEO 212 -OH and for the implementation of, for example, a means that allows antibody coupling of the cross-linkable amphiphilic block copolymer 40 PI O1 -I)-PEO 212 - OH.
  • the functionalisation of the -OH terminal functional group to a carboxyl-terminated group is promising, because the carboxyl-terminated group can be further activated by well known chemical means. Activation of the carboxyl-terminated group by EDC/sulfo-NHS provides the advantage that enables protein coupling for essential techniques.
  • carboxyl groups were introduced to the cross-linkable amphiphilic block copolymer 40 Pl 6 i-b-PEO 212 -OH.
  • the carboxyl groups were introduced in the form of protected ethyl esters during the process of polymerisation.
  • the carboxyl groups were introduced by terminating the polymerisation with ethyl 2-bromoacetate.
  • the resultant ethyl-ester can be hydrolyzed to the carboxyl group by hydrolysis with 0.5 M NaOH at room temperature.
  • the ethyl-ester can be hydrolyzed to the carboxyl group by the enzyme pig liver esterase or the enzyme thermitase.
  • a Fourier transform spectra (see Figure 12) of the cross -linkable amphiphilic block copolymer 40 Pl 61 -b-PE ⁇ 212 -COOH shows a characteristic vibration of the carboxyl group at 1735 cm “1 .
  • the resonance at 1735 cm “1 is typical of H-bridging with the ether functions of the PEO block of the cross-linkable amphiphilic block copolymer 40 Pl ⁇ i-b- PEO 212 -COOH.
  • the methylene stretch resonance of poly (ethylene oxide) and methylene & methyl stretches of the poly (isoprene) appear between 2983 and 2780 cm "1 , while additional deformations of methylene emerge at 1466 and methyl at 1340 cm “1 respectively.
  • the -OH terminal functional group of the cross-linkable amphiphilic block copolymer 40 Pl 6 i-b-PEO 2 i 2 -OH provides a further means to functionalise the -OH to an aldehyde to manufacture the cross -linkable amphiphilic block copolymer 40 Pl ⁇ i-b- PEO 212 -CHO.
  • the -OH terminal functional group was oxidised by Dess-Martin perio- dinane (DMP) to manufacture the Pl 6 i-b-PEO 212 -CHO aldehyde.
  • DMP Dess-Martin perio- dinane
  • the cross-linkable amphiphilic block copolymer 40 PI O1 -ID-PEO 212 -CHO was identified by 1 H-NMR spectros- copy (see Figure 13).
  • the -OH terminal functional group of the cross-linkable amphiphilic block copolymer 40 Pl 6 i-b-PEO 212 -OH provides a further means to functionalise the -OH to an alkyne to manufacture the cross -linkable amphiphilic block copolymer 40 with a terminal alkyne group.
  • the terminal alkyne was introduced by CDI coupling. A characteristic car- bonyl urethane stretch of the alkyne group was observed at 1767 cm "1 in the FT-IR spectrum (see Figure 14).
  • the -OH terminal functional group of the cross-linkable amphiphilic block copolymer 40 Pl 6 i-b-PEO 212 -OH provides a further means to functionalise the -OH to an alkene to manufacture the cross-linkable amphiphilic block copolymer 40 with a terminal alkene group.
  • the terminal alkene was introduced by CDI coupling.
  • the 1 H-NMR spectra of a cross-linkable amphiphilic block copolymer 40, Pl 6 i-b-PEO 212 -ALKENE confirmed the manufacture of the cross-linkable amphiphilic block copolymer 40, PI O1 - b-PEO 212 -ALKENE.
  • the -OH terminal functional group of the cross-linkable amphiphilic block co- polymer 40 Pl 6 i-b-PEO 212 -OH provides a further means to functionalise the -OH terminal functional group to an azide.
  • the azide was further reduced to an amine by the use of triphenylphosphine or lithium aluminium hydride.
  • the triphenylphosphine or lithium aluminium hydride allows the selective reduction of the azide to an amine without reacting with the double bounds of the PI blocks of the cross -linkable amphiphilic block copolymer 40.
  • the -OH terminal functional group was functionalised to an amine by CDI coupling of ethylenediamine (see Figure 17).
  • the terminal amine of the cross -linkable amphiphilic block copolymer 40 Pl 6 i-b-PEO 212 -NH 2 allows further activation steps of the terminal amine by bi-functional homo and hetero molecules like ⁇ , ⁇ sulfo-NHS active esters. Furthermore a reaction of the terminal amine allows for a further functionalisation reaction with aldehydes to form a terminal imine (Schiff base), which can be reduced with sodium cyanoborohydride to a secondary amine.
  • the -OH terminal functional group of the cross-linkable amphiphilic block copolymer 40 Pl 6 i-b-PEO 2 i 2 -OH provides a further means to functionalise the -OH terminal functional group to a terminal boronic acid.
  • the manufacture of the terminal boronic acid ester is achieved by a simple esterification in an aqueous medium.
  • the cross -linkable amphiphilic block copolymer 40 PI O1 -I)-PEO 212 - BORONIC ACID ESTER was identified by 1 H-NMR spectroscopy (see Figure 18).
  • the -OH terminal functional group of the cross -linkable amphiphilic block copolymer 40 Pl 6 i-b-PEO 212 -OH provides a further means to functionalise the -OH terminal functional group to an epoxide with epichlorohydrine.
  • Scavenger-columns can, for example, be placed directly in the solution.
  • the 1,4-regio- isomer of 1,2,3-triazol was identified by 1 H-NMR spectroscopy (see Figure 20).
  • 2,3,4,6-tetra-O-acetyl-l-azido-l-deoxy-/?-D-galactopyranose (GalAc-N3) and 2,3,4,6- tetra-O-acetyl-l-azido-l-deoxy-/?-D-glucopyranose (GluAc-N3) were coupled the cross- linkable amphiphilic block copolymer 40 PI O1 -I)-PEO 212 -OH.
  • the cross-linked micellar nanoparticle complex 50 exhibits benefits for use in biomedical applications.
  • the nanoparticle 5 is a quantum dot.
  • Theoretical modelling studies have shown that two spectral windows are available for in vivo quantum dot imaging at 700 - 900 nm and at 1200 - 1600 nm. The spectral windows are important for deep tissue imaging, because the spectral windows are distinct from the major absorption peaks of blood and water which are found in in vivo applications.
  • the quantum dots are favourable for molecular imaging in contrast to genetically encoded fluorescent proteins, such as green fluorescent protein (GFP), which is also applied for cancer imaging due to the to the 10-50 times larger molar extinction coefficient (0.5 - 0.2 • 10 6 M -1 Cm "1 ) of the quantum dots than those of the organic dyes (5 - 10 • 10 4 M 1 Cm "1 ). Furthermore the quantum dots absorption rates are 10-50 times faster than those of organic dyes at the same excitation photon flux which makes quantum dots appear 10- 20 times brighter than organic dyes.
  • GFP green fluorescent protein
  • quantum dots aggregate non-selectively on the surface of cell membranes (see for example S. Pathak, S. -K. Choi, N. Arnheim, M. E. Thompson, J. Am. Chem. Soc. 2001, 123, 4103).
  • an antibody is attached to the PEO block of the cross -linkable amphiphilic block copolymer 40 PI O1 -I)-PEO 2I2 -OH of the cross-linked micellar nanoparticle complex 50.
  • the protein antibody is attached to the out- ermost area of the cross-linked micellar nanoparticle complex 50, thus conserving the flexibility and accessibility of the antibody to the in vivo environment.
  • the PEO block and a protein-nanoparticle interaction is known not only to minimize non-specific adsorption with proteins in the biological target but also for enhancing the stability of attached antibody molecules (see for example P. Ghosh, G. Han, M. De, C. K. Kim, V. M. Rotello, Adv. Drug Delivery Rev. 2008, 60, 1307).
  • the cross -linkable amphiphilic block copolymer 40 PIe 1 -I)-PEO 212 -OH provides additional negative charges where the cross-linkable amphiphilic block copolymer 40 PI O1 - b-PEO 212 -OH is functionalised with carboxyl groups.
  • the negative charges are known to increase blood circulation time of bio-chemical substances (see for example X. Gao, Y. Cui, R. M. Levenson, L. W. K. Chung, S. Nie, Nat. Biotechnol. 2004, 22, 969).
  • the antibodies and antibody fragments were coupled to the cross -linkable amphiphilic block copolymer 40 PI O1 -I)-PEO 212 -OH of the cross-linked micellar nanoparticle complex 50, under sterile conditions utilising EDC/sulfo-NHS activation.
  • the antibodies murine IgG2b K anti-YKL-40 (146 kD protein), mouse IgG2b isotype (146 kD protein) and DIG-F(ab')2 (100 kD protein) were coupled to the cross -linkable amphiphilic block copolymer 40 Pl 6 i-b-PEO 212 -COOH.
  • the activation of the cross-linkable amphiphilic block copolymer 40 PI O1 -I)-PEO 212 -COOH with EDC/sulfo-NHS proceeds in an analogous man- ner to carbodiimide coupling reactions.
  • the sulfo-NHS activated cross-linkable amphiphilic block copolymer 40 species has enhanced stability at pH ⁇ 6.
  • the cross-linked micellar nanoparticle complex 50 solution was transferred into vessels and subjected with three equivalents of the antibodies (or antibody- fragments). After ten hours of stirring the solution was again dialysed against PBS at pH 7.4 to remove uncoupled antibodies.
  • the final protein coupled cross-linked micellar nanoparticle complex solutions had an antibody concentration of between 0.2 to 0.4 mg/mL.
  • the cross-linked micellar nanoparticle complex 50 that comprises the quantum dot nanoparticle 5 was introduced into a biological target for in vivo analysis.
  • the cross-linked micellar nanoparticle complex 50 comprised the cross -linkable amphiphilic block copolymer 40 functionalised with antibody anti-YKL-40 and the quantum dot nanoparticle 5 (86 pmol) and in a further example the cross-linked micellar nanoparticle complex 50 comprising the cross -linkable amphiphilic block co- polymer 40 functionalised with the protein antibody anti-YKL-40- and a Fe 3 O 4 nanoparticle 5 (300 pmol) conjugate in 100 ⁇ L PBS, pH 7.4, was intravenously injected into a murine tumor model (LOX melanoma cell line)
  • LOX melanoma cell line murine tumor model
  • the protein YKL-40 belongs to the glycosyl hydrolase family 18 and is expressed by several types of tumour (breast, colon, lung, kidney, ovary, prostate, uterine, pancreas, osteosarcoma, thyroid, oligodendroglioma, glioblastoma, and germ cell).
  • tumour breast, colon, lung, kidney, ovary, prostate, uterine, pancreas, osteosarcoma, thyroid, oligodendroglioma, glioblastoma, and germ cell.
  • An iso-control is a control with an antibody such as IgG which can not bind to the tumour specifically.
  • the use of the iso-control demonstrates that there is a specific binding concerning the anti-YKL-40 antibody-quantum dot conjugate. Two mice were applied with the iso-control and 2 mice were applied with the the anti-YKL-40 antibody-quantum dot conjugate.
  • tumour tissue was cut out from both the mice and examined under a confocal microscope. With the detection of the anti-YKL-40-quantum dot (see Figure 23A) the results could be corroborated. Once again the IgG2b-isotype-quantum dot conjugate could not be identified (see Figure 23B), which concludes that the binding of the anti-YKL-40- quantum dot conjugate was not by reason on unspecific binding.
  • a quantum dot probe can also be delivered to the tumour by passive targeting mechanisms. It is known that macromolecules and nanometre-sized parti- cles accumulate preferentially at the tumour through an enhanced permeability and a retention effect (see for example R. K. Jain, J. Controlled Release 2001, 74, 7). This passive targeting was not observed by the present applicants as illustrated in Figure 23B. The non observation of passive targeting of the tumour could be due to the low concentration of the cross-linked micellar nanoparticle complex 50 and/or the excess negative charges of the free carboxylic groups present on the cross-linked micellar nanoparticle complex 50.
  • Negative charges are known to reduce the rate of probe delivery and to subsequent accumulation into tumour tissues (see for example R. B. Campbell, D. Fukumura, E. B. Brown, L. M. Mazzola, Y. Izumi, R. K. Jain, V. P. Torchilin, L. L. Munn, Cancer Research, 2002, 62, 6831).
  • the cross-linked micellar nanoparticle complex 50 which comprises antibodies can also be expanded by the addition of co-molecules such as radioactive markers, dyes, and even drugs.
  • the co-molecules can be coupled to the terminal function of the PEO block of the cross-linked micellar nanoparticle complex 50, if the co-molecules are hydro- philic, or if the co-molecules if they are hydrophobic they can be co-encapsulated in the PI block (see Figure 24).
  • Alexa Fluor® 594 was coupled (see Figure 25) to the cross-linkable amphiphilic block copolymer 40 PI O1 -I)-PEO 2I2 -COOH to allow F ⁇ rster resonance energy transfer (FRET) measurements.
  • the FRET measurements are used to testify the density and length of the cross -linkable amphiphilic block copolymer 40 in an aqueous media.
  • the determination of the density and length of the cross-linkable amphiphilic block copolymer 40 in an aqueous media is valuable for the production of FRET based biosensors (see for example B. Dubertret, M. Calame, A. J. Libchaber, Nature Biotechnology 2001, 19, 365).
  • the Alexa Fluor® 594 coupled cross -linkable amphiphilic block copolymer 40 was identified by 1 H- NMR spectroscopy (see Figure 25).
  • the significance of the cross-linked micellar nanoparticle complex 50 is remarkable.
  • the transfer of the cross-linkable polymer ligand 20 and the cross-linkable amphiphilic block copolymer 40 around a nanoparticle 5, followed by the formation of the intramolecular cross-linkage 60, provides a route for using nanoparticles 5 in biomedical applications.
  • the use of nanoparticle clusters of different ones of the nanoparticles 5 in the cross-linked micellar nanoparticle complex 50 allows a combination of diagnostic and imaging applications of the nanoparticle 5.
  • two nanoparticles 5 of different size are present in the cross-linked micellar nanoparticle complex 50, this would allow fluorescence intensity multiplexing.
  • the functional groups present on the cross-linked mi- cellar nanoparticle complex 50 facilitate the attachment of further chemical groups such as organic fluorophores, radioactive markers, and drugs.
  • CdSe/CdS/ZnS core-shell-shell nanoparticles with a passivation ligand layer of tri-n-octylphosphine oxide (TOPO) and tri-n-octylphosphine (TOP) were synthesised by methods reported previously (I. Mekis, D. V. Talapin, A. Kornowski, M. Haase, H. Weller, Journal of Physical Chemistry B 2003, 107, 7454 and D. V. Talapin, A. L. Rogach, A. Kornowski, M. Haase, H. Weller, Nano Letters 2001, 1, 207).
  • the passivated nanoparticle 10 was precipitated twice with methanol to remove the excess TOPO and was stored in chloroform. Size selective precipitation of the passivated nanoparticles 10 was performed by the stepwise addition of methanol and following centrifugation. Each precipitate was stored separately.
  • Lanthanide phosphate passivated nanoparticles 10 were prepared in an organic solution containing diphenyl ether, phosphoric acid, and dihexyl ether, according to published procedures (O. Lehmann, H. Meyssamy, K. Koempe, H. Schnablegger, M. Haase, J. Phys. Chem. B 2003, 107, 7449). After the diphenyl ether has been removed by vacuum distillation at 100 0 C, tetrahydrofuran (THF) and TOPO was added to the nanopar- tide solution.
  • THF tetrahydrofuran
  • Fe 3 O 4 passivated nanoparticle 10 which were coated with sodium oleate, were obtained by a large scale synthesis (see the method described by J. Park, K. An, Y. Hwang, J.-G. Park, H.-J. Noh, J.-Y. Kim, J.-H. Park, , N.-M. Hwang, T. Hyeon, Nature Materials, 2004, 12, 891) and in high-boiling ether solvents (see the method described by U. I. Tromsdorf, N. C. Bigall, M. G. Kaul, O. T. Brans, M. S. Nikolic, B. Mollwitz, R.A. Sperling, R. Reimer, H. Hohenberg, W. J. Parak, S. Foerster, U. Beisiegel, G. Adam, H. Weller, Nano Letters 2007, 7, 2422.).
  • ethylene oxide (88 g, 2.0 mol) was purified in a three step procedure.
  • the ethylene oxide was dried over CaH 2 , sodium surface and n- butyl lithium under inert conditions.
  • the ethylene oxide was distilled to the solution of THF solution of PI.
  • 9 mL of a solution of diphenylmethylpotassium ( ⁇ 1 mol/L 9 mmol) in cyclohexane was added to the THF solution to deprotonate the terminal hydroxyl group of the PI.
  • the solution was stirred for 72 h at 40 0 C and the polymerisation reaction was terminated by the addition of 4 mL acetic acid (90 mmol).
  • cross-linkable amphiphilic block copolymer 40 polyisoprene-block-poly (ethylene oxide) was purified by precipitation by the addition of cold acetone and was analysed by 1 H- NMR, Maldi-TOF and GPC measurements.
  • a hydroxyl terminated polyisoprene polymer was functionalised with 2,2'- diaminodiethylamine (DETA, -N3) by activation of the hydroxyl group by the addition of a twenty fold excess of l,l'-carbonyldiimidazol (CDI) in anhydrous chloroform. After fourteen hours of stirring at room temperature, the excess of CDI was hydrolysed. The resultant solution was washed twice with water and dried over sodium sulphate. A twenty fold excess of 2,2'-diaminodiethylamine (DETA) was added slowly to the solution. The reaction mixture was stirred for further twelve hours at of 55 0 C.
  • DETA 2,2'- diaminodiethylamine
  • the PI-N3 product was precipi- tated twice in ethanol to remove the CDI and the DETA.
  • the PI-N3 was analysed by x H- NMR spectroscopy with a yield of 100%.
  • the passivated nanoparticles 10 were incubated with a three hundred excess of the PI 30 -N3 ligand and three fold precipitation of the resultant capped micellar nanoparticles 30 with ethanol from THF.
  • the capped micellar nanoparticles 30 were stored in chloroform or toluene.
  • the concentration of the capped nanoparticle 30, chloroform stock solution was determined by UV-Vis absorbance.
  • PI-N3 capped nanoparticles were re-suspended in 100 to 300 ⁇ L of the THF solution, in which a fifty fold to five hundred fold molar excess of the cross linkable amphiphilic block copolymer 40 (PI-b-PEO) and a 50 to 100 hundred molar excess of initiator 70 was present.
  • the incubation time varied from one minute up to four hours.
  • the resultant solution was injected to 1000 ⁇ L of water very slowly, with si- multaneous stirring of the water whilst under a nitrogen flow from above of the water surface area. The nitrogen flow facilitates the removal of the THF solution.
  • the solution was diluted with 25 rnL diethyl ether and 25 rnL ethyl acetate.
  • the aldehyde was purified by precipitation at 4 0 C in diethyl ether.
  • the aldehyde was washed three times with cold diethyl ether.
  • the white solid consisting mainly of sodium azide, which was not soluble in chloroform, was separated by centrifugation and discarded.
  • the colourless solid matter which was obtained by purification, was dissolved in water and lyophilised.
  • a third variant was achieved by CDI activation of PI-b-PEO.
  • 2040 mg PI 61 - ⁇ -PEO 212 -OH (15.11 • 10 "5 mol) was dissolved in 15 mL pure chloroform under inert conditions.
  • the obtained solution was added drop wise under stirring to a solution, comprising 480 mg CDI (14.81 • 10 ⁇ 4 mol, 20 eq) in 10 rnL chloroform.
  • After 24 hours stirring at room temperature the mixture was extracted three times with always 20 mL water to hydrolyse unreacted CDI. Having dried the solution of sodium sulfate and removing the solid phase by filtering, the solvent of the filtrate was evaporated.
  • the reaction mixture was stirred for 43 hours at a temperature of 45 0 C, whereon the solvent was evaporated.
  • the residue was dissolved in 60 mL acetone and precipitated by cooling to a temperature of -15 0 C.
  • the precipitate was again dissolved in 2 mL chloroform and the precipitation was repeated three times in acetone and three times in diethyl ether.
  • the product was finally dissolved in water and lyophylized.
  • the final solution of the cross-linked nanoparticle complex has an antibody concentration between 0.2 to 0.4 mg/mL.
  • micellar nanoparticle 30 - capped micellar nanoparticle
  • micellar nanoparticle complex 50 - cross-linked micellar nanoparticle complex 60 - intramolecular cross linkage 70 - cross -linkage initiator

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Abstract

La présente invention concerne un procédé de fabrication, et les produits de celui-ci, d'un complexe de nanoparticules micellaires réticulés (50). Le complexe de nanoparticules micellaires réticulés (50) comprend une réticulation intramoléculaire (60) entre un copolymère séquencé amphiphile pouvant être réticulé (40) et un ligand polymère pouvant être réticulé (20) qui est lié à une nanoparticule (5). Le complexe de nanoparticules micellaires réticulés (50) stabilise la nanoparticule (5) pour son utilisation dans des applications biomédicales.
PCT/EP2010/052540 2009-03-02 2010-03-01 Procédé de fabrication d'un complexe de nanoparticules micellaires réticulés et produits de celui-ci WO2010100108A1 (fr)

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Cited By (8)

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Publication number Priority date Publication date Assignee Title
WO2012001012A2 (fr) 2010-06-28 2012-01-05 Centrum Für Angewandte Nanotechnologie (Can) Gmbh Combinaison micellaire comprenant une nanoparticule et une pluralité de ligands tensioactifs polymérisables (surfomères)
WO2012154332A2 (fr) * 2011-04-04 2012-11-15 William Marsh Rice University Nanoparticules stables pour conditions hautement salines
DE102014114834A1 (de) 2014-10-13 2016-04-14 Centrum Für Angewandte Nanotechnologie (Can) Gmbh Nanopartikel enthaltende Polymermizellen in nicht-wässriger Lösung, Methoden zu ihrer Herstellung und ihrer Anwendung
US9925303B2 (en) * 2012-11-13 2018-03-27 Edwards Lifesciences Corporation Methods for cross-linking bioprosthetic tissue using bio-orthogonal binding pairs
CN111248224A (zh) * 2020-03-04 2020-06-09 北京科技大学 基于MXene量子点的抗菌剂的制备及抗菌活性测试方法
CN112961666A (zh) * 2021-03-08 2021-06-15 西南石油大学 一种压裂液用纳米复合交联剂及其制备方法
CN113249108A (zh) * 2021-05-14 2021-08-13 西南石油大学 一种耐超高温压裂液及其制备方法
CN114181341A (zh) * 2021-12-30 2022-03-15 广东粤港澳大湾区国家纳米科技创新研究院 一种超声引发制备量子点荧光微球的方法

Citations (8)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US6248847B1 (en) 1997-12-31 2001-06-19 Hyundai Electronics Industries Co., Ltd. Copolymer resin, preparation thereof, and photoresist using the same
US20040115433A1 (en) 2001-05-10 2004-06-17 Abdelhamid Elaissari Composite particles,derived conjugates,preparation method and applications
US20040142578A1 (en) 2002-03-28 2004-07-22 Ulrich Wiesner Thin film nanostructures
WO2005102396A2 (fr) 2004-04-20 2005-11-03 Emory University Nanostructures de multimodalite, procedes de fabrication et d'utilisation associes
WO2007018647A2 (fr) 2005-05-02 2007-02-15 Emory University Nanostructures polyvalentes, procedes permettant de les synthetiser et procedes permettant de les utiliser
WO2008054523A2 (fr) 2006-05-04 2008-05-08 Emory University Nanostructures, procédés de synthèse de celles-ci et procédés d'utilisation de celles-ci
WO2009025623A1 (fr) 2007-08-23 2009-02-26 Agency For Science, Technology And Research Polymérisation à la surface de particules avec des micelles inverses
WO2009061456A2 (fr) 2007-11-07 2009-05-14 University Of Houston Nanoparticules d'oxyde de fer superparamagnétique ultra-petites et leurs utilisations

Patent Citations (8)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US6248847B1 (en) 1997-12-31 2001-06-19 Hyundai Electronics Industries Co., Ltd. Copolymer resin, preparation thereof, and photoresist using the same
US20040115433A1 (en) 2001-05-10 2004-06-17 Abdelhamid Elaissari Composite particles,derived conjugates,preparation method and applications
US20040142578A1 (en) 2002-03-28 2004-07-22 Ulrich Wiesner Thin film nanostructures
WO2005102396A2 (fr) 2004-04-20 2005-11-03 Emory University Nanostructures de multimodalite, procedes de fabrication et d'utilisation associes
WO2007018647A2 (fr) 2005-05-02 2007-02-15 Emory University Nanostructures polyvalentes, procedes permettant de les synthetiser et procedes permettant de les utiliser
WO2008054523A2 (fr) 2006-05-04 2008-05-08 Emory University Nanostructures, procédés de synthèse de celles-ci et procédés d'utilisation de celles-ci
WO2009025623A1 (fr) 2007-08-23 2009-02-26 Agency For Science, Technology And Research Polymérisation à la surface de particules avec des micelles inverses
WO2009061456A2 (fr) 2007-11-07 2009-05-14 University Of Houston Nanoparticules d'oxyde de fer superparamagnétique ultra-petites et leurs utilisations

Non-Patent Citations (43)

* Cited by examiner, † Cited by third party
Title
0. LEHMANN; H. MEYSSAMY; K. KOEMPE; H. SCHNABLEGGER; M. HAASE, J. PHYS. CHEM. B, vol. 107, 2003, pages 7449
B. DUBERTRET; M. CALAME; A. J. LIBCHABER, NATURE BIOTECHNOLOGY, vol. 19, 2001, pages 365
B. DUBERTRET; P. SKOURIDES; D. J. NORRIS; V. NOIREAUX; A. H. BRIVANLOU; A. LIBCHABER, SCIENCE, vol. 298, 2002, pages 1759
B.-S. KIM; J.-M. QIU; J.-P. WANG; T. A. TATON, NANO LETT, vol. 5, 2005, pages 1987
C. A. LEATHERDALE; M. G. BAWENDI, MATERIALS PHYSICS, vol. 63, 2001, pages 165315,1
C. A. LEATHERDALE; M. G. BAWENDI, PHYSICAL REVIEW B: CONDENSED MATTER AND MATERIALS PHYSICS, vol. 63, 2001, pages 165315,1
C. M. NIEMEYER, ANGEW. CHEM., INT. ED., vol. 40, 2001, pages 4128
D. S. LIDKE; P. NAGY; R. HEINTZMANN; D. J. ARNDT-JOVIN; J. N. POST; H. E. GRECCO; E. A. JARES-ERIJMAN; T. M. JOVIN, NAT. BIOTECHNOL, vol. 22, 2004, pages 198
D. V. TALAPIN; A. L. ROGACH; A. KORNOWSKI; M. HAASE; H. WELLER, NANO LETTERS, vol. 1, 2001, pages 207
G. GOPALAKRISHNAN; C. DANELON; P. IZEWSKA; M. PRUMMER; P.-Y. BOLINGER; I. GEISSBUHLER; D. DEMURTAS; J. DUBOCHET; H. VOGEL, ANGEW. CHEM., INT. ED., vol. 45, 2006, pages 478
G. SOLTESZ EDWARD; S. KIM; G. LAURENCE RITA; M. DEGRAND ALEC; P. PARUNGO CHERIE; M. DOR DELPHINE; H. COHN LAWRENCE; G. BAWENDI MOU, THE ANNALS OF THORACIC SURGERY, vol. 79, 2005, pages 269
H. FAN; E. W. LEVE; C. SCULLIN; J. GABALDON; D. TALLANT; S. BUNGE; T. BOYLE; M. C.WILSON; C. J. BRINKER, NANO LETT. NANO LETTERS, vol. 5, 2005, pages 645
H. FAN; E. W. LEVE; C. SCULLIN; J. GABALDON; D. TALLANT; S. BUNGE; T. BOYLE; M. C.WILSON; C. J. BRINKER, NANO LETTERS, vol. 5, 2005, pages 645
H. GU; K. XU; C. XU; B. XU, CHEMICAL COMM., 2006, pages 941
I. MEKIS; D. V. TALAPIN; A. KORNOWSKI; M. HAASE; H. WELLER, JOURNAL OF PHYSICAL CHEMISTRY, vol. 107, 2003, pages 7454
J. A. NEFF; K. D. CALDWELL; P. A. TRESCO, J. BIOMED. MATER. RES., vol. 40, 1998, pages 511
J. K. JAISWAL; H. MATTOUSSI; J. M. MAURO; S. M. SIMON, NAT. BIOTECHNOL. NATURE BIOTECHNOLOGY, vol. 21, 2003, pages 47
J. PARK; K. AN; Y. HWANG; J.-G. PARK; H.-J. NOH; J.-Y. KIM; J.-H. PARK; N.-M. HWANG; T. HYEON, NATURE MATERIALS, vol. 12, 2004, pages 891
J.-H. LEE; Y.-W. JUN; S.-I. YEON; J.-S. SHIN; J. CHEON, ANGEWANDTE CHEMIE, INTERNATIONAL EDITION, vol. 45, 2006, pages 8160
K. MANDAL SWAPAN; N. LEQUEUX; B. ROTENBERG; M. TRAMIER; J. FATTACCIOLI; J. BIBETTE; B. DUBERTRET, LANGMUIR SURFACES AND COLLOIDS, vol. 21, 2005, pages 4175
KANG Y ET AL: "Core/shell gold nanoparticles by self-assembly and crosslinking of micellar, block-copolymer shells", ANGEWANDTE CHEMIE - INTERNATIONAL EDITION 20050103 WILEY-VCH VERLAG DE LNKD- DOI:10.1002/ANIE.200461119, vol. 44, no. 3, 3 January 2005 (2005-01-03), pages 409 - 412, XP002584216 *
M. A. GOLUB; C. L. STEPHENS, J. POLYM. SCI., PART A-1: POLYM. CHEM., vol. 6, 1968, pages 763
M. E. AKERMAN; W. C. W. CHAN; P. LAAKKONEN; S. N. BHATIA; E. RUOSLAHTI, PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE USA, vol. 99, 2002, pages 12617
M. SVENSSON; P. ALEXANDRIDIS; P. LINSE, MACROMOLECULES, vol. 32, 1999, pages 637
NIKOLIC M S ET AL: "Tailor-made ligands for biocompatible nanoparticles", ANGEWANDTE CHEMIE - INTERNATIONAL EDITION 20061006 WILEY-VCH VERLAG DE LNKD- DOI:10.1002/ANIE.200602209, vol. 45, no. 39, 6 October 2006 (2006-10-06), pages 6577 - 6580, XP002584218 *
P. ALIVISA- TOS, NATURE BIOTECHNOLOGY, vol. 22, 2004, pages 47
P. GHOSH; G. HAN; M. DE; C. K. KIM; V. M. ROTELLO, ADV. DRUG DELIVERY REV., vol. 60, 2008, pages 1307
Q. ZHANG; E. E. REMSEN; K. L. WOOLEY, J. AM. CHEM. SOC., vol. 122, 2000, pages 3642
R. B. CAMPBELL; D. FUKUMURA; E. B. BROWN; L. M. MAZZOLA; Y. IZUMI; R. K. JAIN; V. P. TORCHILIN; L. L. MUNN, CANCER RESEARCH, vol. 62, 2002, pages 6831
R. K. JAIN, J. CONTROLLED RELEASE, vol. 74, 2001, pages 7
R. LARSON; W. R. ZIPFEL; R. M. WIL- LIAMS; S. W. CLARK; M. P. BRUCHEZ; F. W. WISE; W. WEBB, SCIENCE, vol. 300, 2003, pages 1434
S. JOHANSEN JULIA; V. JENSEN BENNY; A. ROSLIND; D. NIELSEN; A. PRICE PAUL: "Cancer Epidemiol. Biomarkers & Prev.", vol. 15, 2006, AMERICAN ASSOCIATION FOR CANCER RESEARCH, pages: 194
S. KIM; Y. T. LIM; E. G. SOLTESZ; A. M. DE GRAND; J. LEE; A. NAKAYAMA; J. A. PARKER; T. MIHALJEVIC; R. G. LAURENCE; D. M. DOR, NAT. BIOTECHNOL., vol. 22, 2004, pages 93
S. PATHAK; S.-K. CHOI; N. ARNHEIM; M. E. THOMPSON, J. AM. CHEM. SOC., vol. 123, 2001, pages 4103
T. R. SATHE; A. AGRAWAL; S. NIE, ANAL. CHEM., vol. 78, 2006, pages 5627
U. I. TROMSDORF; N. C. BIGALL; M. G. KAUL; O. T. BRUNS; M. S. NIKOLIC; B. MOLLWITZ; R.A. SPER- LING; R. REIMER; H. HOHENBERG; W. J, NANO LETTERS, vol. 7, 2007, pages 2422
V. BUETUEN; X. S. WANG; M. V. D. P. BANEZ; K. L. ROBINSON; N. C. BILLINGHAM; S. P. ARMES; Z. TUZAR, MACROMOLECULES, vol. 33, 2000, pages 1
W. LIU; M. HOWARTH; A. B. GREYTAK; Y. ZHENG; D. G. NOCERA; A. Y. TING; M. G.BAWENDI, J. AM. CHEM. SOC., vol. 130, 2008, pages 1274
WARREN S C ET AL: "Ordered mesoporous materials from metal nanoparticle-block copolymer self-assembly", SCIENCE AMERICAN ASSOCIATION FOR THE ADVANCEMENT OF SCIENCE USA, vol. 320, no. 5884, 27 June 2008 (2008-06-27), pages 1748 - 1752, XP002584217, ISSN: 0036-8075 *
WOOLEY, BIOMACROMOLECULES, vol. 6, no. 5, 2005, pages 2541 - 2554
X. GAO; Y. CUI; R. M. LEVENSON; L. W. K. CHUNG; S. NIE, NAT. BIOTECHNOL., vol. 22, 2004, pages 969
X. MICHALET; F. F. PINAUD; L. A. BENTOLILA; J. M. TSAY; S. DOOSE; J. J. LI; G. SUNDARESAN; A. M. WU; S. S. GAMBHIR; S. WEISS, SCIENCE, vol. 307, 2005, pages 538
Z. GAO; A. N. LUKYANOV; A. SINGHAL; V. P. TORCHILIN, NANO LETT., vol. 2, 2002, pages 979

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WO2012001012A2 (fr) 2010-06-28 2012-01-05 Centrum Für Angewandte Nanotechnologie (Can) Gmbh Combinaison micellaire comprenant une nanoparticule et une pluralité de ligands tensioactifs polymérisables (surfomères)
WO2012154332A2 (fr) * 2011-04-04 2012-11-15 William Marsh Rice University Nanoparticules stables pour conditions hautement salines
WO2012154332A3 (fr) * 2011-04-04 2014-05-01 William Marsh Rice University Nanoparticules stables pour conditions hautement salines
US9925303B2 (en) * 2012-11-13 2018-03-27 Edwards Lifesciences Corporation Methods for cross-linking bioprosthetic tissue using bio-orthogonal binding pairs
US11213611B2 (en) 2012-11-13 2022-01-04 Edwards Lifesciences Corporation Cross-linked bioprosthetic tissue using bio-orthogonal binding pairs
WO2016059020A2 (fr) 2014-10-13 2016-04-21 Centrum Für Angewandte Nanotechnologie (Can) Gmbh Micelles polymères contenant des nanoparticules dans une solution non aqueuse, leurs procédés de préparation et utilisation
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WO2016059019A2 (fr) 2014-10-13 2016-04-21 Centrum Für Angewandte Nanotechnologie (Can) Gmbh Micelles polymères contenant des nanoparticules en solution non aqueuse, leurs procédé de préparation et d'utilisation
US10315178B2 (en) 2014-10-13 2019-06-11 Förderung Der Angewandten Forschung E.V. Polymer micelles containing nanoparticles in non-aqueous solution, methods for their preparation and use
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DE102014114834A1 (de) 2014-10-13 2016-04-14 Centrum Für Angewandte Nanotechnologie (Can) Gmbh Nanopartikel enthaltende Polymermizellen in nicht-wässriger Lösung, Methoden zu ihrer Herstellung und ihrer Anwendung
CN111248224A (zh) * 2020-03-04 2020-06-09 北京科技大学 基于MXene量子点的抗菌剂的制备及抗菌活性测试方法
CN111248224B (zh) * 2020-03-04 2021-03-16 北京科技大学 基于MXene量子点的抗菌剂的制备及抗菌活性测试方法
CN112961666A (zh) * 2021-03-08 2021-06-15 西南石油大学 一种压裂液用纳米复合交联剂及其制备方法
CN113249108A (zh) * 2021-05-14 2021-08-13 西南石油大学 一种耐超高温压裂液及其制备方法
CN114181341A (zh) * 2021-12-30 2022-03-15 广东粤港澳大湾区国家纳米科技创新研究院 一种超声引发制备量子点荧光微球的方法
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