US20070154965A1 - Chlorotoxin-labeled nanoparticle compositions and methods for targeting primary brain tumors - Google Patents

Chlorotoxin-labeled nanoparticle compositions and methods for targeting primary brain tumors Download PDF

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US20070154965A1
US20070154965A1 US11/600,004 US60000406A US2007154965A1 US 20070154965 A1 US20070154965 A1 US 20070154965A1 US 60000406 A US60000406 A US 60000406A US 2007154965 A1 US2007154965 A1 US 2007154965A1
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chlorotoxin
cells
labeled
particle
binding
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Miqin Zhang
James Olson
Raymond Sze
Richard Ellenbogen
Omid Veiseh
Conroy Sun
Jonathan Gunn
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University of Washington
Fred Hutchinson Cancer Center
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Assigned to WASHINGTON, UNIVERSITY OF reassignment WASHINGTON, UNIVERSITY OF ASSIGNMENT OF ASSIGNORS INTEREST (SEE DOCUMENT FOR DETAILS). Assignors: ZHANG, MIQIN, SZE, RAYMOND, GUNN, JONATHAN WHITNEY, SUN, CONROY, VEISEH, OMID, ELLENBOGEN, RICHARD
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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
    • A61K49/001—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
    • A61K49/0021—Fluorescence in vivo characterised by the fluorescent group, e.g. oligomeric, polymeric or dendritic molecules the fluorescent group being a small organic molecule
    • A61K49/0041—Xanthene dyes, used in vivo, e.g. administered to a mice, e.g. rhodamines, rose Bengal
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61B—DIAGNOSIS; SURGERY; IDENTIFICATION
    • A61B17/00—Surgical instruments, devices or methods, e.g. tourniquets
    • A61B17/22—Implements for squeezing-off ulcers or the like on the inside of inner organs of the body; Implements for scraping-out cavities of body organs, e.g. bones; Calculus removers; Calculus smashing apparatus; Apparatus for removing obstructions in blood vessels, not otherwise provided for
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K47/00—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient
    • A61K47/50—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates
    • A61K47/51—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the non-active ingredient being a modifying agent
    • A61K47/62—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient 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
    • A61K47/64—Drug-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
    • A61K47/6415—Toxins or lectins, e.g. clostridial toxins or Pseudomonas exotoxins
    • 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
    • A61K49/0013—Luminescence
    • A61K49/0017—Fluorescence in vivo
    • A61K49/0019—Fluorescence in vivo characterised by the fluorescent group, e.g. oligomeric, polymeric or dendritic molecules
    • A61K49/0021—Fluorescence in vivo characterised by the fluorescent group, e.g. oligomeric, polymeric or dendritic molecules the fluorescent group being a small organic molecule
    • A61K49/0032—Methine dyes, e.g. cyanine dyes
    • 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
    • A61K49/0013—Luminescence
    • A61K49/0017—Fluorescence in vivo
    • A61K49/005—Fluorescence in vivo characterised by the carrier molecule carrying the fluorescent agent
    • A61K49/0056—Peptides, proteins, polyamino acids
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P35/00—Antineoplastic agents
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K14/00—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
    • C07K14/435—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans
    • C07K14/43504—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans from invertebrates
    • C07K14/43513—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans from invertebrates from arachnidae
    • C07K14/43522—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans from invertebrates from arachnidae from scorpions
    • 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
    • G01N33/53—Immunoassay; Biospecific binding assay; Materials therefor
    • G01N33/574—Immunoassay; Biospecific binding assay; Materials therefor for cancer
    • G01N33/57484—Immunoassay; Biospecific binding assay; Materials therefor for cancer involving compounds serving as markers for tumor, cancer, neoplasia, e.g. cellular determinants, receptors, heat shock/stress proteins, A-protein, oligosaccharides, metabolites
    • G01N33/57492—Immunoassay; Biospecific binding assay; Materials therefor for cancer involving compounds serving as markers for tumor, cancer, neoplasia, e.g. cellular determinants, receptors, heat shock/stress proteins, A-protein, oligosaccharides, metabolites involving compounds localized on the membrane of tumor or cancer cells
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K38/00—Medicinal preparations containing peptides
    • A61K38/16—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
    • 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
    • A61K49/0013—Luminescence
    • A61K49/0017—Fluorescence in vivo
    • 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
    • A61K49/0013—Luminescence
    • A61K49/0017—Fluorescence in vivo
    • A61K49/005—Fluorescence in vivo characterised by the carrier molecule carrying the fluorescent agent
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K51/00—Preparations containing radioactive substances for use in therapy or testing in vivo
    • A61K51/02—Preparations containing radioactive substances for use in therapy or testing in vivo characterised by the carrier, i.e. characterised by the agent or material covalently linked or complexing the radioactive nucleus
    • A61K51/04—Organic compounds
    • A61K51/08—Peptides, e.g. proteins, carriers being peptides, polyamino acids, proteins

Definitions

  • Gliomas are currently the most common and lethal type of primary brain tumor and are one of the leading causes of cancer-related deaths. Treating malignant gliomas remains a daunting challenge due to the difficulty in differentiating between tumor and healthy brain tissue; the rapid growth rate of invasive gliomas; the sensitivity of normal brain tissue to current therapies; the intrinsic cellular resistance of gliomas to drugs; and the blood brain barrier's ability to prevent the passage of substances including drugs and contrast agents.
  • Magnetic resonance imaging MRI
  • Cltx Chlorotoxin
  • MMP-2 membrane-bound matrix metalloproteinase 2
  • a need for a magnetic nanoparticle conjugated with targeting or therapeutic agents for early cancer detection and treatment through real-time assessment of therapeutic and surgical efficacy exists a need for a probe that allows for improved effectiveness of tumor resection in neurosurgery by providing a visual contrast between neoplastic and normal brain tissue.
  • MRI magnetic resonance imaging
  • the present invention seeks to fulfill these needs and provides further related advantages.
  • the invention provides a chlorotoxin-labeled particle comprising:
  • the nanoparticle comprises:
  • each silane moiety is covalently coupled to the surface, wherein at least one terminus of the plurality of silane moieties comprises a chlorotoxin, and wherein the silane moieties comprise a polyalkylene oxide moiety intermediate the surface and the chlorotoxin.
  • the magnetic nanoparticle including the chlorotoxin comprises:
  • each silane moiety is covalently coupled to the core, and wherein at least one silane moiety has the formula:
  • n 1, 2, 3, 4, 5, or 6;
  • n is an integer from about 10 to about 1000;
  • L is a direct bond or a linker
  • T is a chlorotoxin
  • compositions that include the particles of the invention are provided.
  • the composition includes a nanoparticle suitable for administration to a human.
  • the composition can include an acceptable carrier.
  • the invention provides methods for using nanoparticles.
  • the invention provides a method for differentiating neuroectodermal-derived tumor cells from non-neoplastic brain tissue, comprising:
  • the invention provides a method for detecting neuroectodermal-derived tumor cells, comprising:
  • the invention provides a method for detecting a tissue expressing membrane-bound matrix metalloproteinase (MMP-2) protein complex, comprising:
  • MMP-2 membrane-bound matrix metalloproteinase
  • the invention provides a method for determining the location of glioma cells in a patient pre-operatively, intra-operatively, and post-operatively, comprising:
  • composition comprises a pharmaceutically acceptable carrier and an amount of a fluorophore/chlorotoxin-labeled nanoparticle sufficient to image glioma cells in vivo;
  • the invention provides methods for treating a tissue using the nanoparticles.
  • the invention provides a method for treating a glioma in a patient, comprising administering to a patient in need thereof an effective amount of a pharmaceutical composition comprising a chlorotoxin-labeled nanoparticle and a pharmaceutically acceptable carrier.
  • the invention provides a method for treating a neuroectodermal tumor, comprising administering to a patient in need thereof an effective amount of a pharmaceutical composition comprising a chlorotoxin-labeled nanoparticle and a pharmaceutically acceptable carrier.
  • the invention provides a method for treating a tumor expressing matrix metalloproteinase (MMP-2) protein complex, comprising administering to a patient in need thereof an effective amount of a pharmaceutical composition comprising a chlorotoxin-labeled nanoparticle and a pharmaceutically acceptable carrier.
  • MMP-2 matrix metalloproteinase
  • the invention provides a method for inhibiting invasive activity of neoplastic cells, comprising administering to neoplastic cells an effective amount of a pharmaceutical composition comprising a chlorotoxin-labeled nanoparticle and a pharmaceutically acceptable carrier.
  • FIG. 1A is a schematic illustration of the synthesis of a representative chlorotoxin-labeled nanoparticle of the invention (nanoparticle-PEG-Cltx or NPC);
  • FIG. 1B is a TEM image of representative nanoparticles useful in making the chlorotoxin-labeled nanoparticles of the invention.
  • FIG. 1C is a schematic illustration of chlorotoxin-labeled nanoparticle (NPC) binding to a glioma cell;
  • FIGS. 2A-2F illustrate uptake of a representative chlorotoxin-labeled nanoparticle (NPC) of the invention compared to dextran-coated nanoparticles by 9L cells as a function of nanoparticle concentration as assessed by MR imaging ( FIG. 2A ), the transverse relaxation rate R 2 ( FIG. 2B ) and ICP-AES ( FIG. 2C ), nanoparticle uptake as a function of cell incubation time for NPC and dextran-coated nanoparticles evaluated by MR imaging ( FIG. 2D ), transverse relaxation rate R 2 ( FIG. 2E ) and ICP-AES ( FIG.
  • NPC chlorotoxin-labeled nanoparticle
  • FIGS. 4A-4C are images of representative microscopic fields of 9L cell migration across 8 ⁇ m pores of size matrix without chlorotoxin presence ( FIG. 4A ), in presence of 4 ⁇ g/ml free chlorotoxin ( FIG. 4B ), and in presence of 10 ⁇ g Fe/ml of NPC ( FIG. 4C );
  • FIG. 4D is a bar graph representing the relative number of 9L cells that migrated through the pores under the three conditions; in the images, cell nuclei and cytoplasm appear in dark blue and red respectively, and the circles are the pores of the filters;
  • FIG. 5A is a TEM image of representative nanoparticles useful in preparing the chlorotoxin/fluorophore-labeled nanoparticles of the invention.
  • FIG. 5B is an X-ray diffraction pattern of representative nanoparticles useful in preparing the chlorotoxin/fluorophore-labeled nanoparticles of the invention.
  • FIGS. 6A-6D are schematic illustrations for the preparation of representative chlorotoxin/fluorophore-labeled nanoparticles of the invention.
  • FIG. 7A is a confocal fluorescent image of 9L glioma cells cultured with control fluorophore-labeled nanoparticles (NP-Cy5.5);
  • FIG. 7B is a confocal fluorescent image of 9L glioma cells cultured with representative fluorophore-labeled nanoparticles of the invention (NPC-Cy5.5);
  • FIG. 7C is a magnetic resonance phantom image of 9L glioma cells cultured with control fluorophore-labeled nanoparticles (NP-Cy5.5) and representative fluorophore-labeled nanoparticles of the invention (NPC-Cy5.5) embedded in agarose (4.7 T, spin echo pulse sequence, TR 3000 ms, TE 15 ms);
  • FIG. 8A is a confocal fluorescent image of rat cardiomyocyte (rCM) cells cultured with representative chlorotoxin/fluorophore-labeled nanoparticles of the invention (NPC-Cy5.5);
  • FIG. 8B is a confocal fluorescent image of 9L glioma cells cultured with representative chlorotoxin/fluorophore-labeled nanoparticles of the invention (NPC-Cy5.5);
  • FIG. 8C is a magnetic resonance phantom image of 9L glioma and rCM cells cultured with control chlorotoxin-labeled nanoparticles (NPCs) embedded in agarose (4.7 T, spin echo pulse sequence, TR 3000 ms, TE 15 ms); and
  • FIGS. 9A-9C are confocal fluorescent images of 9L glioma cells cultured with representative chlorotoxin/fluorophore-labeled nanoparticles of the invention (NPC-Cy5.5): FIG. 9A , top section of cells; FIG. 9B , middle section of cells; and FIG. 9C , bottom section of cells.
  • the present invention provides chlorotoxin-labeled nanoparticles capable of targeting primary brain tumors, compositions that include the nanoparticles, methods of imaging tissues using the nanoparticles, and methods for treating cells expressing chlorotoxin binding sites using the nanoparticles.
  • the invention provides a chlorotoxin-labeled particle comprising:
  • the particle includes a core having a surface that can be reacted with the silane compounds of the invention.
  • the particles can be core-shell particles in which the core is a material different from the shell.
  • the surface or shell comprises hydroxyl groups that are reactive toward the silane compounds.
  • the core includes a material having magnetic resonance imaging activity.
  • Suitable materials having magnetic resonance imaging activity include metal oxides, such as ferrous oxide, ferric oxide, silicon oxide, polycrystalline silicon oxide, aluminum oxide, germanium oxide, zinc selenide, tin dioxide, titanium dioxide, indium tin oxide, and gadolinium oxide. Mixtures of one or more metal oxide can be used.
  • the core can include non-magnetic materials, such as silicon nitride, stainless steel, titanium, and nickel titanium. Mixtures of one or more non-magnetic materials can also be used.
  • the core of the particles useful in making the particles of the invention have a diameter of from about 2 nm to about 25 nm.
  • the particles of the invention can be nanoparticles having particle diameter of from about 15 nm to about 200 nm.
  • the chlorotoxin of the particles of the invention can be native chlorotoxin, synthetic chlorotoxin, recombinant chlorotoxin, and fragments and variants thereof having chlorotoxin binding activity.
  • the particles of the invention include from about 1 to about 50 chlorotoxins/particle. In one embodiment, the particles include from about 10 to about 50 chlorotoxins/particle. In one embodiment, the particles include about 10 chlorotoxins/particle.
  • the magnetic nanoparticle of the invention includes a chlorotoxin that serves as a targeting moiety that is effective to direct the nanoparticle to cells expressing chlorotoxin binding sites where the nanoparticle is bound.
  • Primary brain tumor cells e.g., neuroectodermal-derived tumor cells and glioma cells
  • the nanoparticle comprises:
  • each silane moiety is covalently coupled to the surface, wherein at least one terminus of the plurality of silane moieties comprises a chlorotoxin, and wherein the silane moieties comprise a polyalkylene oxide moiety intermediate the surface and the chlorotoxin.
  • the particles of the invention include a plurality of silane moieties covalently coupled to the core's surface.
  • the covalently coupled silane moieties provide a layer surrounding the core.
  • the covalently coupled silane moieties provide a monolayer on the core's surface.
  • Each silane moiety includes a polyalkylene oxide moiety.
  • the polyalkylene oxide is a polyethylene oxide.
  • Suitable polyethylene oxides have molecular weights of from about 100 to about 100,000 g/mole.
  • the nanoparticle including a plurality of silane moieties useful for reaction with the chlorotoxin to provide the chlorotoxin-labeled particles of the invention is prepared as described in Zhang et al., J. Am. Chem. Soc. 2004, 126:7206-7211, expressly incorporated herein by reference in its entirety.
  • the magnetic nanoparticle including the chlorotoxin comprises:
  • each silane moiety is covalently coupled to the core, and wherein at least one silane moiety has the formula:
  • n 1, 2, 3, 4, 5, or 6;
  • n is an integer from about 10 to about 1000;
  • L is a direct bond or a linker
  • T is a chlorotoxin
  • the linker may be one or more atoms that link the chlorotoxin to the polyoxyethylene moiety covalently coupled to the core surface.
  • the chlorotoxin-labeled nanoparticles can further include other useful agents.
  • Other useful agents include diagnostic agents.
  • Suitable diagnostic agents include agents that provide for the detection of the nanoparticle by methods other than magnetic resonance imaging.
  • Suitable diagnostic agents include light-emitting compounds (e.g., fluorophores, phosphors, and luminophors).
  • Suitable fluorophores include fluorophores that can be covalently coupled to silane moiety and that emit fluorescence in the visible and near-infrared region of the spectrum.
  • Representative fluorophores include ALEXA FLUOR, AMCA, BODIPY, CASCADE BLUE, CASCADE YELLOW, coumarins, fluoresceins, eosins, erythrosins, rhodamines, OREGON GREEN, PACIFIC BLUE, and TEXAS RED dyes commercially available in suitable reactive forms from Molecular Probes, Inc.
  • cyanine dyes e.g., Cy2, Cy3, Cy3B, Cy3.5, Cy5, Cy5.5, and Cy7
  • CYDYE fluors commercially available in suitable reactive forms from Amersham Pharmacia Biotech (Piscataway, N.J., now GE Healthcare).
  • the chlorotoxin-labeled particle further comprises a fluorescent moiety.
  • the particles of the invention include from about 1 to about 10 fluorescent moieties/particle. In one embodiment, the particles include from about 1 to about 2 fluorescent moieties/particle. In one embodiment, the particles include about 1.5 fluorescent moieties/particle.
  • the fluorescent moiety is selected from red and near infrared emitting fluorescent moieties (i.e., fluorescent moieties having emission maxima greater than about 600 nm). In one embodiment, the fluorescent moiety is a cyanine moiety. In one embodiment, the fluorescent moiety is a Cy5.5 moiety.
  • radiolabels e.g., radio isotopically labeled compounds
  • 125 I, 14 C, and 31 P examples include 125 I, 14 C, and 31 P, among others.
  • a portion of the plurality of silane moieties comprise a diagnostic agent (e.g., a fluorescent agent or fluorophore, a radiolabel).
  • a diagnostic agent e.g., a fluorescent agent or fluorophore, a radiolabel
  • compositions that include the particles of the invention are provided.
  • the composition includes a nanoparticle suitable for administration to a human or an animal subject.
  • the composition can include an acceptable carrier.
  • the composition is a pharmaceutically acceptable composition and includes a pharmaceutically acceptable carrier.
  • carrier refers to a diluent (e.g., saline) to facilitate the delivery of the particles.
  • the invention provides methods for using nanoparticles.
  • the invention provides a method for differentiating neuroectodermal-derived tumor cells from non-neoplastic brain tissue.
  • neuroectodermal-derived tumor cells are differentiated from non-neoplastic brain tissue by:
  • the invention provides a method for detecting neuroectodermal-derived tumor cells.
  • neuroectodermal-derived tumor cells are detected by:
  • the above methods are useful in differentiating and detecting glioma cells.
  • the invention provides a method for detecting a tissue expressing matrix metalloproteinase (MMP-2) protein complex.
  • a tissue expressing metalloproteinase (MMP-2) protein complex is detected by:
  • MMP-2 matrix metalloproteinase
  • MMP-2 matrix metalloproteinase
  • measuring the level of binding of the chlorotoxin-labeled nanoparticle comprises magnetic resonance imaging.
  • the chlorotoxin-labeled nanoparticle further comprises a fluorescent moiety.
  • measuring the level of binding of the chlorotoxin-labeled nanoparticle can include fluorescence imaging.
  • the invention provides a method for determining the location of glioma cells in a patient pre-operatively, intra-operatively, and post-operatively.
  • the methods includes the steps of:
  • composition comprises a pharmaceutically acceptable carrier and an amount of a fluorophore/chlorotoxin-labeled nanoparticle sufficient to image glioma cells in vivo;
  • an amount of a fluorophore/chlorotoxin-labeled nanoparticle sufficient to image glioma cells in vivo is an amount from about 1-20 mg Fe/kg body weight (“Fe” refers to iron present in particle core.
  • steps (d) and (e) may be repeated.
  • the above method includes pre-operative, intra-operative, and post-operative imaging. It will be appreciated that variations of the above method are within the scope of the invention. Other variations of the method include, for example, (1) pre-operative imaging only; (2) intra-operative imaging only; (3) post-operative imaging only; (4) pre-operative and intra-operative imaging only; (5) pre-operative and post-operative imaging only; and (6) intra-operative and post-operative imaging only.
  • the invention provides methods for treating a tissue using the nanoparticles.
  • the invention provides a method for treating a glioma in a patient, comprising administering to a patient in need thereof an effective amount of a pharmaceutical composition comprising a chlorotoxin-labeled nanoparticle and a pharmaceutically acceptable carrier.
  • the invention provides a method for treating a neuroectodermal tumor, comprising administering to a patient in need thereof an effective amount of a pharmaceutical composition comprising a chlorotoxin-labeled nanoparticle and a pharmaceutically acceptable carrier.
  • the invention provides a method for treating a tumor expressing matrix metalloproteinase (MMP-2) protein complex, comprising administering to a patient in need thereof an effective amount of a pharmaceutical composition comprising a chlorotoxin-labeled nanoparticle and a pharmaceutically acceptable carrier.
  • MMP-2 matrix metalloproteinase
  • the invention provides a method for inhibiting invasive activity of neoplastic cells, comprising administering to neoplastic cells an effective amount of a pharmaceutical composition comprising a chlorotoxin-labeled nanoparticle and a pharmaceutically acceptable carrier.
  • the chlorotoxin-labeled nanoparticles of the invention can be used to detect and treat various cancers (e.g., prostate cancer, sarcomas, hematological malignancies, and leukemias) and various neuroectodermal tumors (e.g., glioma, meningioma, ependymonas, medulloblastoma, neuroblastoma, glioblastoma, ganglioma, pheochromocytoma, melanoma, Ewing's sarcoma, small cell lung carcinoma, and metastatic brain tumors.)
  • cancers e.g., prostate cancer, sarcomas, hematological malignancies, and leukemias
  • various neuroectodermal tumors e.g., glioma, meningioma, ependymonas, medulloblastoma, neuroblastoma, glio
  • malignant cancers include gliomas, astrocytomas medulloblastomas, choroids plexus carcinomas, ependymomas, other brain tumors, neuroblastoma, head and neck cancer, lung cancer, breast cancer, intestinal cancer, pancreatic cancer, liver cancer, kidney cancer, sarcomas (over 30 types), osteosarcoma, rhabdomyosarcoma, Ewing's sarcoma, carcinomas, melanomas, ovarian cancer, cervical cancer, lymphoma, thyroid cancer, anal cancer, colo-rectal cancer, endometrial cancer, germ cell tumors, laryngeal cancer, multiple myeloma, prostate cancer, retinoblastoma, gastric cancer, testicular cancer, and Wilm's
  • the methods of the invention are applicable to human and animal subjects.
  • the present invention provides a chlorotoxin-labeled nanoparticle capable of targeting glioma cells detectable by magnetic resonance (MR) imaging.
  • the nanoparticle can be used to correlate pre-operative and post-operative diagnostic images.
  • the nanoparticle was synthesized by coating iron oxide nanoparticles with covalently-bound bifunctional polyethylene glycol (PEG) that were subsequently functionalized with chlorotoxin.
  • PEG polyethylene glycol
  • the chlorotoxin-labeled nanoparticles of the invention include a polyoxyethylene moiety intermediate the particle and chlorotoxin.
  • the polyoxyethylene moiety is formed covalently coupling a polyethylene glycol to the nanoparticle via self-assembly.
  • the use of the polyoxyethylene linker (PEG) improves cellular internalization, prevents nanoparticle agglomeration, increases blood circulation time in vivo, and offers binding sites for chlorotoxin and fluorescent label.
  • the siloxane bond coupling the nanoparticle with the short PEG chains confers the stability of the PEG self-assembled monolayers (SAMS) and increases PEG packing density on nanoparticles by establishing covalent bonds between PEG interchains.
  • SAMS PEG self-assembled monolayers
  • PEG-coated nanoparticles used in this study had an overall size less than 15 nm as shown in FIG. 1 , which, in combination with the particle's surface chemistry, may offer chlorotoxin-labeled nanoparticles (NPCs) the ability to penetrate the blood brain barrier.
  • NPCs chlorotoxin-labeled nanoparticles
  • Chlorotoxin's small size can facilitate internalization of NPCs, as opposed to widely used antibodies, which are bulky and thus have difficulty crossing the cell membrane.
  • Chlorotoxin was covalently linked to the PEG through a thioether bond foamed between SATA and SIA. This linkage is highly stable under the reducing environments of blood and liver.
  • the delineation of the tumor margins with targeting-magnetic nanoparticles plays a crucial role in the detection and characterization of lesions in situ to guide treatment and determine surgical resection effectiveness.
  • dextran-coated nanoparticles are being studied for intravascular administration in intraoperative MRI examinations to determine and remove tumors with a high degree of certainty.
  • tumor margins are inferred by labeling macrophages situated at the tumor margins, and consequently detected by identifying adjacent microglial cells and not the tumor cells directly. This limits its application for detecting highly invasive brain tumors such as gliomas whose cells may quickly infiltrate surrounding healthy tissue.
  • chlorotoxin limits the invasive nature of tumor cells, lending a therapeutic facet to the NPC system. It has been demonstrated that chlorotoxin inhibits the upregulation of MMP-2, further decreasing the invasive nature of the glioma cells.
  • the matrigel invasion assay confirmed that the bioactivity of chlorotoxin molecules was retained after their conjugation to nanoparticles via the chemical scheme introduced herein. Interestingly, the same therapeutic effect was observed with a lower concentration of chlorotoxin on NPCs versus free chlorotoxin.
  • this therapeutic enhancement is that the PEG on nanoparticles facilitated cellular uptake of NPC and the nanoparticles helped localize the short peptides in the target cells. Limiting tumor cell invasion can prevent further metastasis of the tumor cells to healthy brain tissue and improve surgical outcome.
  • the chlorotoxin-labeled nanoparticle of the invention is a magnetic nanoparticle that serves as both an MRI contrast enhancement agent and a drug carrier to inhibit brain tumor cell invasion. A high degree of internalization by targeted glioma cells was observed making the probe ideally suited for further modification with cytotoxic agents.
  • the chlorotoxin-labeled nanoparticle of the invention demonstrated a strong affinity for gliomas, but not healthy brain cells.
  • the high sensitivity of the nanoparticle of the invention provides an effective approach to the early detection, real-time monitoring, and treatment of brain tumors.
  • FIG. 1C illustrates conceptually the binding of a NPC to a glioma cell, in which the NPC inhibits the activity of MMP-2 endopeptidase and its further expression by glioma cells.
  • the cellular invasion pathway by endopeptidase is blocked.
  • this interaction induces a second physiological response through down regulation of further MMP-2 expression. This dual action alters the invasive behavior of glioma cells.
  • the present invention provides a chlorotoxin-labeled nanoparticle for targeting gliomas and inhibiting tumor migration.
  • the chlorotoxin-labeled nanoparticle is an iron oxide superparamagnetic nanoparticle coated with a chlorotoxin.
  • MRI magnetic resonance imaging
  • ICP-AES inductively coupled plasma atomic emission spectroscopy
  • the specificity of glioma-targeting was evaluated through a comparison of the nanoparticle binding to glioma cells that express matrix metalloproteinase 2 (MMP-2) versus healthy cells that do not express detectable MMP-2.
  • MMP-2 matrix metalloproteinase 2
  • the ability of the nanoparticle to inhibit cell migration was demonstrated by the matrigel invasion test in which the chlorotoxin-labeled nanoparticle showed similar efficacy to free chlorotoxin at a concentration of 4 times lower than that of free chlorotoxin peptide. This indicates that chlorotoxin retains its bioactivity after conjugation to the nanoparticle and that the nanoparticle enhanced the cellular uptake and retention of chlorotoxin in the target cells.
  • the chlorotoxin-labeled nanoparticle can be used for highly sensitive detection of glioma tumors and targeted therapy in the central nervous system.
  • the present invention provides a multifunctional nanoparticle that includes a chlorotoxin and a fluorescent moiety.
  • the chlorotoxin/fluorophore-labeled nanoparticle is capable of targeting glioma cells, detectable by both magnetic resonance imaging and fluorescence imaging. Significant preferential uptake of the chlorotoxin/fluorophore-labeled nanoparticle by glioma cells was identified by both MRI and fluorescence imaging.
  • This multifunctional nanoprobe can be used to image resections of glioma brain tumors in real time and to correlate preoperative diagnostic images with intraoperative pathology at cellular-level resolution.
  • the probe was fabricated by coating iron oxide nanoparticles (NPs) with covalently-bound bifunctional polyethylene glycol (PEG) polymers that were subsequently functionalized with (a) a chlorotoxin (Cltx), a glioma tumor targeting molecule, and (b) a near infrared fluorescing (NIRF) molecule.
  • NPs iron oxide nanoparticles
  • PEG polyethylene glycol
  • Cltx is a unique peptide shown to specifically target the vast majority of glioma tumors.
  • Cltx is a small 36-amino acid peptide purified from the venom of the giant Israeli scorpion ( Leiurus quinquestriatus ). This peptide has been shown to bind with high affinity to the membrane-bound matrix metalloproteinase-2 (MMP-2) endopeptidase, which is preferentially upregulated in gliomas, medulloblastomas, and other tumors of the neuroectodermal origin.
  • MMP-2 membrane-bound matrix metalloproteinase-2
  • NIRF neoplasm originating from healthy brain tissue and allows visualization of tissues millimeters in depth due to the efficient penetration of photons in the near infrared range.
  • PEG coatings were used to prevent nanoparticles from agglomeration and protein adsorption, and thus would increase particle blood circulation time and the efficiency of their internalization by targeted cells when introduced in vivo.
  • BBB blood brain barrier
  • the nanoparticle of the invention Compared to dextran-coated nanoparticles currently used in MRI intraoperative examination for glioma resection, which label macrophages situated at the tumor boundary rather than the tumor cells themselves, the nanoparticle of the invention, by virtue of its covalently coupled chlorotoxin, directly targets tumor cells and thus, can potentially “follow” the cell migration to delineate the tumor boundary in real time. This is particularly beneficial for monitoring high-grade glioma cells that are highly invasive and quickly infiltrate the surrounding healthy tissue.
  • the present invention provides iron oxide nanoparticles (NPs) that have been conjugated with a chlorotoxin (Cltx) and a fluorescent compound (e.g., Cy.5.5) to create a multifunctional nanoprobe that specifically targets glioma cells and that is detectable both magnetically and optically.
  • MRI and confocal fluorescence analysis show strong preferential uptake of the nanoparticles of the invention (e.g., NPC-Cy5.5) by glioma cells over control nanoparticles.
  • a significantly higher degree of internalization of the nanoparticles of the invention by glioma cells versus control cells was observed indicating the preferential targeting abilities of the nanoparticles for gliomas.
  • the high stability and prolonged retention (at least 24 hrs) of the nanoparticles of the invention within targeted cells as demonstrated by confocal imaging are particularly advantageous in intraoperative imaging applications as compared to conventional optical fluorophores conjugated to targeting biomolecules.
  • the cellular-level resolution provided by the nanoparticles of the invention provide accurate delineation of otherwise poorly defined glioma interfaces resulting from their highly invasive morphology.
  • the application of the nanoparticles of the invention for preoperative and postoperative diagnostic imaging with MRI and real-time intraoperative visualization of tumor margins with optical devices is a novel approach that will improve the effectiveness of diagnostic and therapeutic modalities available for brain tumor patients.
  • NPC nanoparticle-PEG-chlorotoxin conjugates
  • succinimidyl iodoacetate Molecular Bioscience, Boulder, CO
  • DMSO dimethyl sulfoxide
  • Excess SIA was removed from the suspension through gel filtration chromatography (PD 10 desalting columns, Amersham Biosciences, Uppsala, Sweden), equilibrated with 20 mM sodium citrate, 0.15 M NaCl buffer (pH 8.0).
  • the resulting thiol of the SATA conjugated-Cltx was deprotected by addition of 30 ⁇ l of a solution containing 25 mM hydroxylamine and 10 mM EDTA and maintained for 1 hr at room temperature.
  • Sulfhydryl-modified Cltx was added to the iodoacetate derivatized nanoparticle solution at a molecular ratio of 50 to 1 and mixed on a shaker overnight at 4° C. Unbound Cltx was removed from the suspension through gel filtration chromatography.
  • Dextran-coated NPs were synthesized as described in Molday et al., J. Immunol. Methods 1982, 52:353-367, expressly incorporated herein by reference in its entirety.
  • FIG. 1B shows an image of the nanoparticles prepared as described above.
  • the nanoparticles are well dispersed and uniform in shape and size.
  • Statistical analysis of the TEM micrographs yielded a nanoparticle size of 10.5 ⁇ 1.5 nm.
  • Rat gliosarcoma (9L, ATCC, Manassas, Va.) and rat cardiomyocytes (rCM, Cell Applications, San Diego, Calif.) cells were grown in DMEM and rCM medium respectively, supplemented with 1% streptomycin/penicillin and 10% fetal bovine serum FBS.
  • rCM rat cardiomyocytes
  • Nanoparticle uptake by cells was determined through the measurement of iron concentrations by inductively coupled plasma atomic emission spectroscopy (ICP-AES).
  • ICP-AES inductively coupled plasma atomic emission spectroscopy
  • Cell samples were prepared by dissolving the cell pellets in 200 ⁇ l of concentrated hydrochloric acid (HC1) for 1 hr at 60° C. and then analyzed on a Jarrell Ash 955 ICP-AES spectrophotometer.
  • FIG. 2A shows an MR phantom image of 9L cells incubated with various concentrations of NPCs and dextran-coated NPs.
  • SI signal intensity
  • FIG. 2B shows a linear increase in the transverse relaxation rate R 2 for 9L cells cultured with NPCs as the concentration of NPC increased and only a slight increase in R 2 for 9L cells cultured with dextran-coated nanoparticles.
  • Significant specificity of NPC's role in targeting 9L cells was further demonstrated by quantification of NPC uptake by 9L cells.
  • a linear increase in NPC uptake was observed as NPC concentration increased from 10 to 100 ⁇ g Fe/ml ( FIG. 2C ). Even at the lowest concentration (10 ⁇ g Fe/ml), NPCs internalized by 9L cells was 10.6 times greater than dextran-coated nanoparticles.
  • the cellular uptake of NPC was 50.0 times higher than dextran-coated nanoparticles.
  • the uptake level of dextran-coated nanoparticles by 9L cells is comparable to the data reported on the uptake of dextran crosslinked iron oxide nanoparticles by 9L cells described in Moore et al., Jmri - Journal Of Magnetic Resonance Imaging 1997; 7:1140-1145.
  • Phantom samples were prepared by suspending 10 6 cells in 50 ⁇ l of 1% low-melting agarose (BioRad, Hercules, Calif.). Cell suspensions were then loaded into a pre-fabricated 12-well agarose sample holder and allowed to solidify at 4° C.
  • MR images were acquired using a 4.7-T Varian MR Spectrometer (Varian, Inc., Palo Alto, Calif.) and a Bruker magnet (Bruker Medical Systems, Düsseldorf, Germany) equipped with a 5 cm volume coil. A spin-echo multisection pulse sequence was selected.
  • T 2 values were obtained using a built-in Varian macro, “t2” fit program, to generate a T 2 map of the acquired images. Then, the transverse relaxation rate R 2 or 1/T 2 values were calculated using the obtained T 2 values.
  • FIG. 2D MR images of 9L cells incubated with fixed nanoparticle concentrations over time (over a time period of 1-4 hrs) are shown in FIG. 2D .
  • 9L cells incubated with NPCs showed strong decrease in SI over time, while the cells incubated with dextran-coated nanoparticles showed little change in SI.
  • the cells incubated with NPC had a tR 2 substantially higher than those incubated with dextran-coated NPs for all time points ( FIG. 2E ).
  • the MRI results correlate well with quantitative data obtained through ICP-AES from the same samples shown in FIG. 2F where uptake of NPCs by 9L cells increased significantly (doubled in 4 hrs) with the incubation time as opposed to the virtually no change in uptake of dextran-coated NPs over time.
  • FIG. 3A illustrates a quantitative comparison of NPCs uptake by 9L cells versus rCM cells, and indicates a 14 fold higher uptake of NPCs by 9L cells than rCM cells.
  • FIG. 1C illustrates conceptually the binding of a NPC to a glioma cell, in which the NPC inhibits the activity of MMP-2 endopeptidase and its further expression by glioma cells.
  • the cellular invasion pathway by endopeptidase is blocked.
  • this interaction induces a second physiological response through down regulation of further MMP-2 expression. This dual action alters the invasive behavior of glioma cells.
  • Transwell migration assay was performed by a method reported previously to assess the bioactivity of chlorotoxin. See Deshane et al., J. Biol. Chem. 2003, 278:4135-4144, expressly incorporated herein in its entirety. Prior to the experiment, matrigel invasion chamber inserts (BD Biosciences, Bedford, Mass.) of 8 ⁇ m pore size were rehydrated. The lower surfaces of the chamber inserts were immersed in 5 ⁇ g/ml solution of vitronectin for 8 hours. 9L cells were then plated at a density of 2.5 ⁇ 10 5 cells per chamber. Cells were treated with solutions of 4 ⁇ g/ml of chlorotoxin, and 10 ⁇ g Fe/ml of NPCs, respectively.
  • the chambers were then incubated in a 37° C. humidified incubator maintained at 5% CO 2 for 24 hrs.
  • the cells on the upper inserts were scrubbed off, and the invaded cells were fixed and stained with Diff-Quik stain kit (IMEB INC., Chicago, Ill.).
  • the filters were then detached from the inserts and mounted on glass slides using immersion oil and imaged on a Nikon E800 upright microscope. Relative inhibition of cell invasion was determined through reduction of migrating cells in presence of chlorotoxin and NPCs. Triplicates of each sample were prepared and cell counts were obtained from five random spots on each filter. The data was then averaged and presented as a relative number of invasive cells per treatment.
  • FIGS. 4A-4C show the optical images of 9L cells after migration through the 8 ⁇ m pores of the matrix for the cells treated with no chlorotoxin, 4 ⁇ g/ml free chlorotoxin, and 10 Fe/ml of NPC ( ⁇ 1 ⁇ g/ml chlorotoxin), respectively.
  • NPC free chlorotoxin
  • Iron oxide (Fe 3 O 4 ) nanoparticles were synthesized via a co-precipitation process of iron chlorides and sodium hydroxide. A 1.5 M sodium hydroxide solution was added dropwise to a deoxygenated solution of iron chloride with a Fe(II)/Fe(III) molar ratio of 0.5 under mechanical stirring and ultrasonication. The precipitation of nanoparticles occurred at pH of 12. The resulting black precipitate was then isolated with a rare-earth magnet and washed with deionized water until a pH of 10.5 was reached.
  • FIG. 5A shows a representative image of the prepared nanoparticles.
  • the nanoparticles are well dispersed and uniform in shape and size.
  • Statistical analysis of the TEM micrograph yielded a nanoparticle size of 10.5 ⁇ 1.5 nm.
  • the X-ray diffraction spectrum of the nanoparticles is shown in FIG.
  • FIGS. 6A-6D A schematic illustration of the preparation is shown in FIGS. 6A-6D .
  • FIG. 6A depicts the overall process consisting of three major steps with each step.
  • FIGS. 6B-6D detail the individual steps.
  • nanoparticles prepared as described above, were first modified with trifluoroethylester terminal PEG silane which was then converted to an amine-terminated PEG silane. See Zhang et al., J. Am. Chem. Soc. 126:7206-7211, 2004.
  • monofunctional N-hydroxysuccinimide (NHS) esters of Cy5.5 were then utilized to covalently couple Cy5.5 to the PEG-coated nanoparticles through reaction with the terminal amine.
  • the remaining terminal amines of the PEG coated nanoparticles were conjugated with chlorotoxin, as shown in FIG. 6D , by (1) reacting the amines of PEG coated nanoparticles with a heterobifunctional linker, succinimidyl iodoacetate (SIA), (2) modifying chlorotoxin with heterobifunctional linker, N-succinimidyl-S-acetylthioacetate (SATA) to render sulfhydryl groups, and (3) conjugating iodoacetate derivatized nanoparticles with sulfhydryl modified chlorotoxin.
  • SIA succinimidyl iodoacetate
  • SATA N-succinimidyl-S-acetylthioacetate
  • chlorotoxin is covalently coupled to the nanoparticle through a stable thioether bond that is not susceptible to cleavage even under harsh environment.
  • succinimidyl iodoacetate Molecular Biosciences, Boulder, Colo.
  • DMSO dimethyl sulfoxide
  • chlorotoxin To functionalize chlorotoxin with sulfhydryl groups, a stock solution of chlorotoxin was prepared by dissolving 200 mg chlorotoxin in 200 ⁇ l of 50 mM bicarbonate buffer at pH 8.5. A solution of SATA (Molecular Bioscience, Boulder, Colo.) in anhydrous DMSO was prepared at a concentration of 1 mg/ml. Eight ⁇ l of the SATA solution was then added to the stock chlorotoxin solution, and the mixture was allowed to react for 3-4 hrs at 4° C.
  • SATA Molecular Bioscience, Boulder, Colo.
  • the resulting sulfhydryl modified chlorotoxin was then added to the iodoacetate modified particles at a molecular ratio of 50 to 1 and shaken overnight in an ice bath. Unreacted chlorotoxin was removed from the suspension through gel filtration chromatography using PD10desalting columns equilibrated with 20 mM sodium citrate, 0.15 M NaCl buffer at pH 8.0.
  • the degree of Cy5.5 labeling of NPs was controlled through stoichiometry and reaction conditions, and quantified by fluorescence spectroscopy. Emission intensity of a dilute sample of nanoparticle-Cy5.5 (50 ⁇ g of Fe/ml) at 689 nm was compared to a linear standard prepared using various concentrations of Cy5.5. The number of NPs was calculated on the assumption that each 10 nm NP (as determined from TEM analysis, FIG. 5A ) had a volume of 5.236 ⁇ 10 ⁇ 25 m 3 and a density of 5.2 kg/m 3 based on the determined Fe 3 O 4 crystal structure as identified by X-ray diffraction ( FIG. 5B ). Using this information the mass of a NP was determined to be 2.728 ⁇ 10 ⁇ 18 g, and the reaction yielded 1.22 fluorophores per NP.
  • the number of chlorotoxin peptides linked to each NP was quantified using the bicinchoninic acid (BCA) assay (Pierce Biotechnology, Rockford, Ill.), and NP concentrations were determined by inductive couple plasma atomic emission spectroscopy. From this analysis, the average number of Cltx molecules per NP was determined to be 10.2.
  • BCA bicinchoninic acid
  • Rat cardiomyocytes (rCM, Cell Applications, San Diego, Calif.) were grown in rat cardiomyocyte cell culture media.
  • 9L glioma cells (American Type Culture Collection, Manassas, Va.) were grown in DMEM medium formulation with high glucose supplemented with sodium pyruvate, 1% streptomycin/penicillin and 10% FBS (Invitrogen, Carlsbad, Calif.). Trypan Blue staining was used to determine cell density and viability, and cell counts were obtained using a hemocytometer.
  • DAPI 6-diamidino-2-phenyindole
  • MR phantom imaging samples were prepared by suspending 10 6 cells in 50 ⁇ l of 1% low-melting agarose (BioRad, Hercules, Calif.). Cell suspensions were loaded into a pre-fabricated 12-well agarose sample holder and allowed to solidify at 4° C. MR imaging was performed using a 4.7-T Bruker imager (Bruker Medical Systems, Düsseldorf, Germany) equipped with a 5 cm volume coil. A spin-echo multisection pulse sequence was selected to acquire MR phantom images. Repetition time (TR) of 3000 msec and variable echo times (TE) of 15-90 msec were used.
  • TR Repetition time
  • TE variable echo times
  • the spatial resolution parameters were as follows: an acquisition matrix of 256 ⁇ 128, field of view of 4 ⁇ 4 cm, section thickness of 1 mm, and 2 averages. Regions of interest (ROIs) of 5.0 mm in diameter were placed in the center of each sample image to obtain signal intensity measurements using NIH ImageJ. T2 values were obtained using VnmrJ “t2” fit program to generate a T2 map of the acquired images.
  • ROIs Regions of interest
  • 9L rat glioma cells were cultured with NP-Cy5.5 and NPC-Cy5.5, and their fluorescence confocal images are shown in FIGS. 7A and 7B , respectively.
  • the cellular membrane, nuclei, and NPC-Cy5.5 are green, blue and red, respectively.
  • 9L cells cultured with NPC-Cy5.5 ( FIG. 7B ) took up a substantial amount of NPC-Cy5.5 as clearly identified by infrared (IR) signals, while those cultured with NP-Cy5.5 ( FIG. 7A ) took up virtually no NP-Cy5.5.
  • FIG. 7C shows an MR phantom image of 9L cells cultured with NPCs (top) and control NPs (bottom), respectively.
  • 9L cells cultured with NPCs show a much greater negative contrast than the cells cultured with the control NPs.
  • the corresponding T2 relaxation time of 9L cells with NPCs and control NPs were 5 ms and 95 ms, respectively.
  • FIGS. 8A and 8B show the confocal fluorescence images of rCM and 9L cells, respectively, cultured with NPC-Cy5.5. The images show that the 9L cells have taken up notably higher amounts of NPC-Cy5.5 than the rCM cells. The nanoparticles appeared to be in cytoplasm surround the nuclei.
  • 8C shows a MR phantom image of 9L (top) and rCM (bottom) cells incubated with NPCs.
  • the MRI results are consistent with those obtained through confocal imaging: rCM cells were barely detectable from the agarose background while 9L cells showed dramatically preferential uptake of NPC versus rCM cells.
  • the corresponding T2 relaxation of the 9L cells and rCM cells incubated with NPCs were 15.3 ⁇ 1.7 ms and 63.8 ⁇ 2.2 ms, respectively.
  • FIGS. 9A-9C shows images from three sectional depths of 9L cells. Illustrated from left to right are the corresponding top ( FIG. 9A ), middle ( FIG. 9B ), and bottom ( FIG. 9C ) sections of the cells, with the strongest Cy5.5 fluorescence observed in the middle section, i.e., within the cells and with decreasing fluorescence intensity towards the top and bottom sections.

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Cited By (14)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US20050142062A1 (en) * 1995-12-27 2005-06-30 Sontheimer Harald W. Novel method of diagnosing and treating gliomas
US20080279780A1 (en) * 2005-04-22 2008-11-13 Washington, University Of Fluorescent chlorotoxin conjugate and method for intra-operative visualization of cancer
US20090098574A1 (en) * 2006-04-25 2009-04-16 Centre National De La Recherche Scientifique (Cnrs) Functionalization of gold nanoparticles with oriented proteins, application to the high-density labelling of cell membranes
WO2009126835A2 (fr) * 2008-04-09 2009-10-15 University Of Washington Techtransfer Invention Licensing Nanoparticules magnétiques, et procédé d'imagerie de cellules t
US20100210546A1 (en) * 2002-05-31 2010-08-19 Transmolecular, Inc. Combination chemotherapy with chlorotoxin
US20100285496A1 (en) * 2007-07-30 2010-11-11 University Of Maryland Baltimore Methods and Devices for the Detection of Biofilm
US20110171123A1 (en) * 2008-09-02 2011-07-14 University Of Maryland, Baltimore In Vivo Biofilm Infection Diagnosis and Treatment
WO2011142858A2 (fr) 2010-05-11 2011-11-17 Fred Hutchinson Cancer Research Center Variants de la chlorotoxine, conjugués, et leurs méthodes d'utilisation
US9018347B2 (en) 2010-02-04 2015-04-28 Morphotek, Inc. Chlorotoxin polypeptides and conjugates and uses thereof
US9023595B2 (en) 2008-05-15 2015-05-05 Morphotek, Inc. Treatment of metastatic tumors
US9784730B2 (en) 2013-03-21 2017-10-10 University Of Washington Through Its Center For Commercialization Nanoparticle for targeting brain tumors and delivery of O6-benzylguanine
US10156559B2 (en) 2012-12-10 2018-12-18 Fred Hutchinson Cancer Research Center Lipocalin fusion partners
US10555911B2 (en) 2012-05-04 2020-02-11 Yale University Highly penetrative nanocarriers for treatment of CNS disease
US11559580B1 (en) 2013-09-17 2023-01-24 Blaze Bioscience, Inc. Tissue-homing peptide conjugates and methods of use thereof

Families Citing this family (10)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US9283288B2 (en) 2006-02-24 2016-03-15 Medibeacon, Inc. Methods of using optical agents
US8529874B2 (en) * 2007-05-16 2013-09-10 Ge Healthcare As Peptide imaging agents
JP2010535811A (ja) * 2007-08-07 2010-11-25 トランスモレキュラー， インコーポレイテッド 薬物キャリアとしてのクロロトキシン
EP2211913A4 (fr) * 2007-10-12 2010-12-22 Transmolecular Inc Administration systémique d'agents de chlorotoxine pour le diagnostic et le traitement de tumeurs
US7892760B2 (en) * 2007-11-19 2011-02-22 Celera Corporation Lung cancer markers, and uses thereof
JP5497006B2 (ja) * 2008-04-14 2014-05-21 ザ ジェネラル ホスピタル コーポレイション 膵管腺癌の検出および治療のためのプレクチン−１標的化剤
WO2013003507A1 (fr) * 2011-06-27 2013-01-03 Morphotek, Inc. Agents multifonction
BR112016005802A2 (pt) * 2013-09-17 2017-09-12 Blaze Bioscience Inc conjugados de clorotoxina e métodos de seu uso
WO2017180789A2 (fr) 2016-04-12 2017-10-19 Blaze Bioscience, Inc. Méthodes de traitement faisant intervenir des conjugués de chlorotoxine
US20190282661A1 (en) 2016-04-15 2019-09-19 Blaze Bioscience, Inc. Methods of treating breast cancer

Citations (9)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US5905027A (en) * 1995-12-27 1999-05-18 Uab Research Foundation Method of diagnosing and treating gliomas
US6130101A (en) * 1997-09-23 2000-10-10 Molecular Probes, Inc. Sulfonated xanthene derivatives
US6514481B1 (en) * 1999-11-22 2003-02-04 The Research Foundation Of State University Of New York Magnetic nanoparticles for selective therapy
US20030201208A1 (en) * 2000-04-26 2003-10-30 Martin Koch Dynamic superparamagnetic markers
US20040101822A1 (en) * 2002-11-26 2004-05-27 Ulrich Wiesner Fluorescent silica-based nanoparticles
US20040105980A1 (en) * 2002-11-25 2004-06-03 Sudarshan Tirumalai S. Multifunctional particulate material, fluid, and composition
US6767635B1 (en) * 1999-09-14 2004-07-27 Biomedical Apherese Systeme Gmbh Magnetic nanoparticles having biochemical activity, method for the production thereof and their use
US6972326B2 (en) * 2001-12-03 2005-12-06 Molecular Probes, Inc. Labeling of immobilized proteins using dipyrrometheneboron difluoride dyes
US7462446B2 (en) * 2005-03-18 2008-12-09 University Of Washington Magnetic nanoparticle compositions and methods

Family Cites Families (35)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
DE3407582A1 (de) 1984-03-01 1985-09-05 Dornier System Gmbh, 7990 Friedrichshafen Schaltungsanordnung fuer einen regelkreis
US6667156B2 (en) 1995-12-27 2003-12-23 Uab Research Foundation Diagnosis and treatment of neuroectodermal tumors
US6703381B1 (en) 1998-08-14 2004-03-09 Nobex Corporation Methods for delivery therapeutic compounds across the blood-brain barrier
WO2003000203A2 (fr) 2001-06-26 2003-01-03 Uab Research Foundation Inhibition par la chlorotoxine de l'invasion cellulaire, des metastases cancereuses, de l'angiogenese et du remodelage des tissus
US20060088899A1 (en) 2002-05-31 2006-04-27 Alvarez Vernon L Combination chemotherapy with chlorotoxin
MXPA04011871A (es) 2002-05-31 2005-07-26 Transmolecular Inc Quimioterapia en combinacion con clorotoxina.
DE10328251A1 (de) 2003-06-24 2005-01-13 Toximed Gmbh Pharmazeutischer Wirkstoff
WO2005053611A2 (fr) 2003-11-26 2005-06-16 Transmolecular, Inc. Traitement de troubles phospholipidiques phosphatidylinositols
JP2007532879A (ja) 2004-04-06 2007-11-15 トランスモレキュラー， インコーポレイテッド 骨髄およびリンパ系細胞癌の診断および処置
US20080153746A1 (en) 2004-04-06 2008-06-26 Transmolecular, Inc. Diagnosis and Treatment of Myeloid and Lymphoid Cell Cancers
GB0422901D0 (en) 2004-10-14 2004-11-17 Ares Trading Sa Lipocalin protein
GB0504767D0 (en) 2005-03-08 2005-04-13 Ares Trading Sa Lipocalin protein
EP1877428B1 (fr) 2005-04-22 2011-03-16 University of Washington Conjugue fluorescent de cyanine et procede de visualisation intra-operative de cancer
CN102703446B (zh) 2005-08-18 2014-05-28 Ambrx公司 tRNA组合物和其用途
US20070237714A1 (en) 2006-03-31 2007-10-11 Alvarez Vernon L Diagnosis and treatment of tumors
EP2029157A4 (fr) 2006-05-19 2009-11-18 Georgia Tech Res Inst Ligand des transporteurs abc
CN101003788A (zh) 2006-09-21 2007-07-25 武汉大学 一种蝎抗肿瘤转移肽及其制备方法和应用
CN100564517C (zh) 2006-09-21 2009-12-02 武汉大学 一种蝎抗神经胶质瘤肽及其制备方法和应用
CA2667678A1 (fr) 2006-10-25 2008-07-24 Amgen Inc. Agents therapeutiques a base de peptides toxiques
US20090004105A1 (en) 2007-06-27 2009-01-01 Zhen Cheng Molecular imaging of matrix metalloproteinase expression using labeled chlorotoxin
JP2010535811A (ja) 2007-08-07 2010-11-25 トランスモレキュラー， インコーポレイテッド 薬物キャリアとしてのクロロトキシン
EP2211913A4 (fr) 2007-10-12 2010-12-22 Transmolecular Inc Administration systémique d'agents de chlorotoxine pour le diagnostic et le traitement de tumeurs
AU2008352925A1 (en) 2008-03-20 2009-09-24 Transmolecular, Inc. Inhibition of angiogenesis
CN101270158B (zh) 2008-04-30 2010-12-22 武汉大学 一种靶向抗神经胶质瘤蛋白及制备方法和用途
CA2724384A1 (fr) 2008-05-15 2009-11-19 Transmolecular, Inc. Traitement de tumeurs metastatiques
EP2313430B1 (fr) 2008-06-24 2018-05-02 Technische Universität München Mutéines de hngal et protéines associées ayant une affinité pour une cible donnée
US20100098637A1 (en) 2008-09-23 2010-04-22 The Regents Of The University Of Michigan Dye-loaded nanoparticle
CN101381405B (zh) 2008-09-24 2011-07-27 武汉摩尔生物科技有限公司 基因工程肿瘤靶向kct-w1多肽及制备方法和用途
EP3483610A1 (fr) 2009-11-09 2019-05-15 University Of Washington Center For Commercialization Points de polymère chromophore fonctionnalisé et leurs bioconjugués
CN101921769B (zh) 2010-01-11 2012-07-25 山西大学 一种重组腺病毒及其制备方法和应用
WO2011094671A2 (fr) 2010-01-29 2011-08-04 The Uab Research Foundation Polypeptides à conjugaison n-terminale pour la thérapie et le diagnostic ciblés
CN106995493B (zh) 2010-02-04 2021-09-24 卫材公司 氯毒素多肽和结合物及其应用
CN101824084A (zh) 2010-04-08 2010-09-08 山西大学 靶向性抗胶质瘤蛋白质及其应用
WO2011142858A2 (fr) 2010-05-11 2011-11-17 Fred Hutchinson Cancer Research Center Variants de la chlorotoxine, conjugués, et leurs méthodes d'utilisation
CN107082803A (zh) 2010-08-16 2017-08-22 皮里斯制药有限公司 铁调素的结合蛋白

Patent Citations (9)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US5905027A (en) * 1995-12-27 1999-05-18 Uab Research Foundation Method of diagnosing and treating gliomas
US6130101A (en) * 1997-09-23 2000-10-10 Molecular Probes, Inc. Sulfonated xanthene derivatives
US6767635B1 (en) * 1999-09-14 2004-07-27 Biomedical Apherese Systeme Gmbh Magnetic nanoparticles having biochemical activity, method for the production thereof and their use
US6514481B1 (en) * 1999-11-22 2003-02-04 The Research Foundation Of State University Of New York Magnetic nanoparticles for selective therapy
US20030201208A1 (en) * 2000-04-26 2003-10-30 Martin Koch Dynamic superparamagnetic markers
US6972326B2 (en) * 2001-12-03 2005-12-06 Molecular Probes, Inc. Labeling of immobilized proteins using dipyrrometheneboron difluoride dyes
US20040105980A1 (en) * 2002-11-25 2004-06-03 Sudarshan Tirumalai S. Multifunctional particulate material, fluid, and composition
US20040101822A1 (en) * 2002-11-26 2004-05-27 Ulrich Wiesner Fluorescent silica-based nanoparticles
US7462446B2 (en) * 2005-03-18 2008-12-09 University Of Washington Magnetic nanoparticle compositions and methods

Cited By (28)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US20050142062A1 (en) * 1995-12-27 2005-06-30 Sontheimer Harald W. Novel method of diagnosing and treating gliomas
US20100210546A1 (en) * 2002-05-31 2010-08-19 Transmolecular, Inc. Combination chemotherapy with chlorotoxin
US8778310B2 (en) 2005-04-22 2014-07-15 University Of Washington Fluorescent chlorotoxin conjugate and method for intra-operative visualization of cancer
US20080279780A1 (en) * 2005-04-22 2008-11-13 Washington, University Of Fluorescent chlorotoxin conjugate and method for intra-operative visualization of cancer
US20090098574A1 (en) * 2006-04-25 2009-04-16 Centre National De La Recherche Scientifique (Cnrs) Functionalization of gold nanoparticles with oriented proteins, application to the high-density labelling of cell membranes
US20130217037A1 (en) * 2006-04-25 2013-08-22 Alain Brisson Functionalization of gold nanoparticles with oriented proteins, application to the high-density labeling of cell membranes
US20100285496A1 (en) * 2007-07-30 2010-11-11 University Of Maryland Baltimore Methods and Devices for the Detection of Biofilm
US8541006B2 (en) 2007-07-30 2013-09-24 University Of Maryland, Baltimore Methods and devices for the detection of biofilm
WO2009126835A2 (fr) * 2008-04-09 2009-10-15 University Of Washington Techtransfer Invention Licensing Nanoparticules magnétiques, et procédé d'imagerie de cellules t
WO2009126835A3 (fr) * 2008-04-09 2010-03-04 University Of Washington Techtransfer Invention Licensing Nanoparticules magnétiques, et procédé d'imagerie de cellules t
US9603952B2 (en) 2008-05-15 2017-03-28 Morphotek, Inc. Treatment of metastatic tumors
US9023595B2 (en) 2008-05-15 2015-05-05 Morphotek, Inc. Treatment of metastatic tumors
US20110171123A1 (en) * 2008-09-02 2011-07-14 University Of Maryland, Baltimore In Vivo Biofilm Infection Diagnosis and Treatment
US8697375B2 (en) * 2008-09-02 2014-04-15 University Of Maryland, Baltimore In vivo biofilm infection diagnosis and treatment
US9637526B2 (en) 2010-02-04 2017-05-02 Morphotek, Inc. Chlorotoxin polypeptides and conjugates and uses thereof
US10183975B2 (en) 2010-02-04 2019-01-22 Morphotek, Inc. Chlorotoxin polypeptides and conjugates and uses thereof
US9018347B2 (en) 2010-02-04 2015-04-28 Morphotek, Inc. Chlorotoxin polypeptides and conjugates and uses thereof
US9234015B2 (en) 2010-02-04 2016-01-12 Morphotek, Inc. Chlorotoxin polypeptides and conjugates and uses thereof
US20130195760A1 (en) * 2010-05-11 2013-08-01 Fred Hutchinson Cancer Research Center Chlorotoxin variants, conjugates, and methods for their use
WO2011142858A2 (fr) 2010-05-11 2011-11-17 Fred Hutchinson Cancer Research Center Variants de la chlorotoxine, conjugués, et leurs méthodes d'utilisation
US9944683B2 (en) * 2010-05-11 2018-04-17 Fred Hutchinson Cancer Research Center Chlorotoxin variants, conjugates, and methods for their use
KR20130113938A (ko) * 2010-05-11 2013-10-16 프레드 헛친슨 켄서 리서치 센터 클로로톡신 변이체, 접합체, 및 이의 사용 방법
KR101972173B1 (ko) 2010-05-11 2019-04-24 프레드 헛친슨 켄서 리서치 센터 클로로톡신 변이체, 접합체, 및 이의 사용 방법
US10822381B2 (en) 2010-05-11 2020-11-03 Fred Hutchinson Cancer Research Center Chlorotoxin variants, conjugates, and methods for their use
US10555911B2 (en) 2012-05-04 2020-02-11 Yale University Highly penetrative nanocarriers for treatment of CNS disease
US10156559B2 (en) 2012-12-10 2018-12-18 Fred Hutchinson Cancer Research Center Lipocalin fusion partners
US9784730B2 (en) 2013-03-21 2017-10-10 University Of Washington Through Its Center For Commercialization Nanoparticle for targeting brain tumors and delivery of O6-benzylguanine
US11559580B1 (en) 2013-09-17 2023-01-24 Blaze Bioscience, Inc. Tissue-homing peptide conjugates and methods of use thereof

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