WO2004096190A1 - Composition comprenant une nanoparticule magnetique enveloppant un materiau magnetique et un medicament a l'aide d'un polymere synthetique biodegradable - Google Patents

Composition comprenant une nanoparticule magnetique enveloppant un materiau magnetique et un medicament a l'aide d'un polymere synthetique biodegradable Download PDF

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WO2004096190A1
WO2004096190A1 PCT/KR2004/001024 KR2004001024W WO2004096190A1 WO 2004096190 A1 WO2004096190 A1 WO 2004096190A1 KR 2004001024 W KR2004001024 W KR 2004001024W WO 2004096190 A1 WO2004096190 A1 WO 2004096190A1
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magnetic nanoparticles
drug
magnetic
set forth
synthetic polymer
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PCT/KR2004/001024
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Si-Young Song
Seung-Joo Ham
Seong-Bae Park
Jin-Gu Gang
Han-Soo Kim
Yong-Gun Shul
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Yonsei University
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    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K9/00—Medicinal preparations characterised by special physical form
    • A61K9/48—Preparations in capsules, e.g. of gelatin, of chocolate
    • A61K9/50—Microcapsules having a gas, liquid or semi-solid filling; Solid microparticles or pellets surrounded by a distinct coating layer, e.g. coated microspheres, coated drug crystals
    • A61K9/51—Nanocapsules; Nanoparticles
    • A61K9/5107—Excipients; Inactive ingredients
    • A61K9/5115—Inorganic compounds
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K9/00—Medicinal preparations characterised by special physical form
    • A61K9/48—Preparations in capsules, e.g. of gelatin, of chocolate
    • A61K9/50—Microcapsules having a gas, liquid or semi-solid filling; Solid microparticles or pellets surrounded by a distinct coating layer, e.g. coated microspheres, coated drug crystals
    • A61K9/51—Nanocapsules; Nanoparticles
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K41/00—Medicinal preparations obtained by treating materials with wave energy or particle radiation ; Therapies using these preparations
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K9/00—Medicinal preparations characterised by special physical form
    • A61K9/0002—Galenical forms characterised by the drug release technique; Application systems commanded by energy
    • A61K9/0009—Galenical forms characterised by the drug release technique; Application systems commanded by energy involving or responsive to electricity, magnetism or acoustic waves; Galenical aspects of sonophoresis, iontophoresis, electroporation or electroosmosis
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K9/00—Medicinal preparations characterised by special physical form
    • A61K9/48—Preparations in capsules, e.g. of gelatin, of chocolate
    • A61K9/50—Microcapsules having a gas, liquid or semi-solid filling; Solid microparticles or pellets surrounded by a distinct coating layer, e.g. coated microspheres, coated drug crystals
    • A61K9/51—Nanocapsules; Nanoparticles
    • A61K9/5107—Excipients; Inactive ingredients
    • A61K9/513—Organic macromolecular compounds; Dendrimers
    • A61K9/5138—Organic macromolecular compounds; Dendrimers obtained by reactions only involving carbon-to-carbon unsaturated bonds, e.g. polyvinyl pyrrolidone, poly(meth)acrylates
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K9/00—Medicinal preparations characterised by special physical form
    • A61K9/48—Preparations in capsules, e.g. of gelatin, of chocolate
    • A61K9/50—Microcapsules having a gas, liquid or semi-solid filling; Solid microparticles or pellets surrounded by a distinct coating layer, e.g. coated microspheres, coated drug crystals
    • A61K9/51—Nanocapsules; Nanoparticles
    • A61K9/5107—Excipients; Inactive ingredients
    • A61K9/513—Organic macromolecular compounds; Dendrimers
    • A61K9/5146—Organic macromolecular compounds; Dendrimers obtained otherwise than by reactions only involving carbon-to-carbon unsaturated bonds, e.g. polyethylene glycol, polyamines, polyanhydrides
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K9/00—Medicinal preparations characterised by special physical form
    • A61K9/48—Preparations in capsules, e.g. of gelatin, of chocolate
    • A61K9/50—Microcapsules having a gas, liquid or semi-solid filling; Solid microparticles or pellets surrounded by a distinct coating layer, e.g. coated microspheres, coated drug crystals
    • A61K9/51—Nanocapsules; Nanoparticles
    • A61K9/5192—Processes
    • 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

Definitions

  • the present invention relates, in general, to a composition applicable in a targeted drug delivery system, and, more particularly, to a composition including magnetic nanoparticles each of which encapsulates a magnetic material and a drug within biodegradable synthetic polymers, a method of preparing the magnetic nanoparticles, and a targeted drug delivery system capable of concentrating the magnetic nanoparticles to a target site using a magnetic field.
  • the targeted drug delivery system used in research and clinical applications includes one of the following basic schemes: (a) direct application of a drug to an affected site (organ or tissue); (b) passive accumulation of a drug via the leaky vasculature, for example, at the site of tumor, infarction or inflammation; (c) physical targeting at a target site such as a tumor or inflammation site based on abnormal pH and/or temperature (e.g., using a drug carrier sensitive to pH and temperature); (d) targeting of a drug attached to a magnetic carrier by the action of an external l magnetic field; and (e) employment of a vector molecule with a highly specific affinity to an affected site.
  • U.S. Patent No. 4,345,588 discloses a method of delivering a therapeutic agent to a target capillary bed of the body, which is characterized by preparing intravascularly-administrable, magnetically-localizable biodegradable microspheres containing a therapeutic agent and administering for permanently localization in the target capillary bed for release of the therapeutic agent therein.
  • the particles disclosed in the above U.S. patent are limited in their application due to their micron size and thus incapable of penetrating the intact intracellular space.
  • the microparticles are encapsulated with natural substances such as lipids, proteins or carbohydrates, their isolation and washing are not easily accomplished.
  • the present invention provides a composition including magnetic nanoparticles which are capable of substantially concentrating a drug at a target site and deeply penetrating the target site to improve bioavailability of the drug and minimize the undesirable side effects of the drug.
  • the present invention provides a composition including magnetic nanoparticles each of which encapsulates a magnetic material and a drug with a biodegradable synthetic polymer.
  • the present invention provides a method of preparing magnetic nanoparticles, comprising the steps of: (a) dissolving a drug, a biodegradable synthetic polymer and a magnetic material in a partially water-soluble solvent (organic phase); (b) saturating the resulting organic solution to an aqueous solution (aqueous phase) of a stabilizing agent to reach equilibrium; (c) emulsifying the resulting saturated solution using a homogenizer; (d) adding water to the resulting emulsified solution to diffuse the partially water-soluble solvent into the aqueous phase; and (e) isolating magnetic nanoparticles from the resulting solution.
  • the present invention provides a method of preparing magnetic nanoparticles, comprising the steps of: (a) dissolving a drug, a magnetic material and a first emulsifier in distilled water to provide a first aqueous phase; (b) dissolving a biodegradable synthetic polymer in a partially water-soluble solvent to provide an organic phase; (c) adding the first aqueous phase to the organic phase and mixing the resulting mixture with agitation to provide a primary W/O type emulsion; (d) adding to the primary W/O type emulsion distilled water (second aqueous phase)in which a second emulsifier has been dissolved and mixing the resulting mixture with agitation to provide a W/O/W type double emulsion; and (e) isolating magnetic nanoparticles from the W/O/W type double emulsion.
  • FIG. 1 shows SEM images of magnetic nanoparticles prepared by a method of preparing magnetic nanoparticles based on an emulsification-difiusion method with an organic phase:aqueous phase ratio of 1 : 1.5 (a), 1 :2 (b), 1 :3 (c) and 1 :4 (d);
  • FIG. 2 shows SEM images of magnetic nanoparticles prepared by the method of preparing magnetic nanoparticles based on the emulsification-diffiision method with a magnetite:PCL ratio of 1:5 (a) and 2:5 (b);
  • FIG. 3 is a graph showing average diameters of magnetic nanoparticles according to the ratio of magnetite to PCL, which are prepared by the method of preparing magnetic nanoparticles based on the emulsification-diffiision method;
  • FIG. 4 shows TEM images of magnetic nanoparticles prepared by the method of preparing magnetic nanoparticles based on the emulsification-diffiision method with a magnetite:PCL ratio of 1 :0 (a), 1 :5 (b), 2:5 (c) and 3 :5 (d);
  • FIG. 5 shows SEM images of magnetic nanoparticles prepared by a method of preparing magnetic nanoparticles based on a multiple-emulsification method using polyvinylalcohol for a second aqueous phase in a concentration of 0.25% (a), 0.5% (b), 1% (c) and 2% (d);
  • FIG. 6 shows SEM images of magnetic nanoparticles prepared by the method of preparing magnetic nanoparticles based on the multiple-emulsification method with a magnetite:PCL ratio of
  • FIG. 7 is a graph showing average diameters of magnetic nanoparticles according to the ratio of magnetite to PCL, which are prepared by the method of preparing magnetic nanoparticles based on the multiple-emulsification method;
  • FIG. 8 shows results of FTTR analysis of magnetite (a), pure PCL particles (b), magnetic nanoparticles (c) prepared based on the emulsification-diffiision method, and magnetic nanoparticles (d) prepared based on the multiple-emulsification method;
  • FIG. 11 shows release patterns of a drug (cisplastin) encapsulated in magnetic nanoparticles (a) prepared based on the emulsification-diffiision method, and magnetic nanoparticles (b) prepared based on the multiple-emulsification method;
  • FIG. 12 is a photograph showing mice as a control group ((a) and (b)) administered with a composition according to the present invention and mice as a test group ((c) and (d)) administered with the composition according to the present invention and then applied with a magnetic field at a left tumor,
  • FIG. 13 shows results of blue iron staining of tumor tissues not applied with a magnetic field, which has been obtained from mice of the test group
  • FIG. 14 shows results of blue iron staining of tumor tissues applied with a magnetic field, which has been obtained from mice of the test group
  • FIG. 15 shows results of blue iron staining of the spleen (a), the pancreas (b) and the liver (c), which all are not tumor tissues, which all have been obtained from mice of the test group; and
  • FIG. 16 is a graph showing in vivo anti-tumor effect of magnetic nanoparticles containing gemcitabin.
  • the present invention provides a composition including magnetic nanoparticles each of which encapsulates a magnetic material and a drug with a biodegradable synthetic polymer.
  • the biodegradable synthetic polymer is selected from polyesters including polylactide, polyl(lactide-co- glycolide), polycaprolactone and polyanhydroride, which are easily biodegraded by serum esterase.
  • PCL polycaprolactone
  • PCL polycaprolactone
  • PCL is slowly degraded and does not form an acidic environment such as polylactide or polyOactide-co-glycolide) . It is degraded to a non-toxic, low molecular weight byproduct accompanied by simultaneous drug release by biodegradation thereof. Therefore, most preferred is PCL as the biodegradable synthetic polymer in the preparation of the magnetic nanoparticles according to the present invention.
  • Examples of the magnetic material useful in the preparation of the magnetic nanoparticles according to the present invention include ferromagnetic compounds. Most preferred is magnetite
  • the magnetic material is in the form of being finely divided into a hyperfine size of less than 100 nm, preferably less than 30 nm, and most preferably less than 10 nm.
  • Such very small sized magnetite may be prepared by a technique known in the art, for example, fine milling, vapor deposition, chemical precipitation, etc. Fine milling in ball mill may be used for preparation of colloidal suspension of magnetite.
  • magnetite fine powder or suspension is commercially available, for example, from the Ferrofluidics Corporation (Burlington, Massachusetts, U.S. A), which ranges from 10 to 20 nm in particle size.
  • the drug capable of being loaded into the magnetic nanoparticle of the present invention does not have any particular limitation in its kind and chemical properties, and may vary depending on diseases to be treated.
  • the drug may be loaded into the magnetic nanoparticle in an aqueous form if being water-soluble or in the form of being dissolved in an organic solvent if being lipid- soluble. Therefore, the magnetic nanoparticles of the present invention may be used for delivery of various water-soluble drugs and hydrophobic drugs to target sites.
  • the composition including the magnetic nanoparticles according to the present invention is capable of concentrating drugs to desired sites, it is useful for delivery of drugs with restricted clinical applications due to their side effects, such as anti-tumor drugs, immunosuppressors or anti- inflammatory drugs.
  • anti-tumor agents useful in the present invention include cisplatin, carboplatin, procarbazine, mechlorethamine, cyclophosphamide, ifosfamide, melphalan, chlorambucil, busulfan, nitrosourea, diactinomycin, daunorubicin, doxorubicin, bleomycin, plicomycin, etoposide, tamoxifen, taxol, taxotere, transplatinum, vincristin, vinblastin and irinotecan.
  • Gemcitabine and cisplatin used as the drug in an aspect of the present invention are used for treating small-cell lung cancer, metastatic ovarian tumor, pancreatic cancer, advanced bladder cancer, and other cancers.
  • these drugs are known to have severe side effects to the kidney and the gastrointestinal system, and thus limited in their clinical applications.
  • immunosuppressive drugs useful in the present invention include cyclophosphamide, azathiopurine, 6-mercaptopurin (6-MP), cytarabine, bromodeoxyuridine (BudR), fluorodeoxyuridine (FudR), methotrexate, mytomycin C, actinomycin D, cortisone, predonisolone, and dexamethasone.
  • anti-inflammatory drugs useful in the present invention include naproxen, diclofenac, indomethacine, sulindac, piroxicam, ibuprofen, azapropazon, nabumeton, tiaprofenic acid, indoprofen, fenoprofen, flurbiprofen, pirazolac, zaltoprofen, nabumetone, bromfenac, ampiroxicam, and lomoxicam.
  • the magnetic nanoparticles according to the present invention may be prepared by a technique well known in the art.
  • the magnetic nanoparticles are prepared using an emulsification-diffiision technique or a multiple-emulsification technique.
  • the diameter of the magnetic nanoparticle is typically smaller than 1,000 nm, but, in the present invention, its diameter is preferably smaller than 500 nm. Therefore the object of the present invention is to prepare magnetic nanoparticles each of which has a diameter of smaller than 500 nm and a uniform size distribution.
  • the present invention provides a method of preparing magnetic nanoparticles based on an emulsification-diffiision technique, comprising the steps of: (a) dissolving a drug, a biodegradable synthetic polymer and a magnetic material in a partially water-soluble solvent (organic phase); (b) saturating the resulting organic solution to an aqueous solution (aqueous phase) with a stabilizing agent to reach equilibrium; (c) emulsifying the resulting saturated solution using a homogenizer; (d) adding water to the resulting emulsified solution to diffuse the partially water- soluble solvent into the aqueous phase; and (e) isolating magnetic nanoparticles from the resulting solution.
  • the magnetic nanoparticles may be isolated by a process including filtration of the solution obtained at step (d), serum replacement, centrifugation and vacuum drying.
  • the term "partially water-soluble solven ' is intended to mean a solvent having both a polar group and a non-polar group such as propylenecarbonate, ethylacetate, benzylalcohol and methylethylketone. Among them, ethylacetate is particularly preferable because of having mild toxicity, proper solubility and low boiling point.
  • sodium lauryl sulfate, polyvinylalcohol, didodecyldimethyl ammonium bromide, and others, which all have low toxicity to the body are useful for the stabilizing agent.
  • the stabilizing agent is preferably dissolved in the aqueous phase in a concentration ranging from about 1% to about 10% (w/v).
  • the magnetic nanoparticles prepared by the above method may vary in morphology and size according to the ratio of the organic phase to the aqueous phase.
  • the aqueous phase is preferably prepared in a volume two to four times as much as the organic phase. If the aqueous phase is prepared in a twofold smaller volume, the obtained particles are not changed in size but present at an aggregated state. In contrast, if the volume of the aqueous phase exceeds four times the volume of the organic phase, the prepared particles have defects on their surface and are not uniform in size and morphology.
  • the ratio of the organic phase to the aqueous phase is about 1 :2, the particles prepared have a relatively small uniform size distribution. Therefore, in the above method of preparing magnetic nanoparticles according to the present invention, the ratio of the organic phase to the aqueous phase is preferably about 1 :2.
  • the particles prepared by the above method may vary in morphology and size according to the ratio of the magnetic material to the biodegradable synthetic polymer.
  • the magnetic material is preferably used in an amount 0.2 to 0.8 times as much as the biodegradable synthetic polymer.
  • the magnetic nanoparticles prepared are slightly increased in size but greatly improved in encapsulation efficiency into the biodegradable synthetic polymer.
  • the magnetic material is most preferably used in an amount 0.8 times as much as the biodegradable synthetic polymer.
  • the present inventors found that, in case of using a hydrophobic drug, this drug is encapsulated with high efficiency by employing the emulsification-diffiision technique.
  • the emulsification-diffiision technique is preferably employed.
  • the present inventors found that the magnetic nanoparticles prepared by the emulsification-diffiision technique have a relatively rapid drug release rate.
  • the present invention provides a method of preparing magnetic nanoparticles based on a multiple-emulsification technique, comprising the steps of: (a) dissolving a drug, a magnetic material and a first emulsifier in distilled water to provide a first aqueous phase; (b) dissolving a biodegradable synthetic polymer in a partially water-soluble solvent to provide an organic phase; (c) adding the first aqueous phase to the organic phase and mixing the resulting mixture with agitation to provide a primary W/O type emulsion; (d) adding to the primary W/O type emulsion distilled water (second aqueous phase)in which a second emulsifier has been dissolved and mixing the resulting mixture with agitation to provide a W/O/W type double emulsion; and (e) isolating magnetic nanoparticles from the W/O/W type double emulsion.
  • the magnetic nanoparticles may be isolated by a process including filtration of the solution obtained at step (d), serum replacement, centrifugation and vacuum drying.
  • the term "partially water-soluble solvent” is intended to mean a solvent having both a polar group and a non-polar group, such as propylenecarbonate, ethylacetate, benzylalcohol and methylethylketone. Among them, ethylacetate is particularly preferable because of having mild toxicity, proper solubility and low boiling point.
  • the first and second emulsifiers are individually selected from among sodium lauryl sulfate, polyvinylalcohol, didodecyldimethyl ammonium bromide, and others, which all have low toxicity to the body.
  • the magnetic nanoparticles prepared by the above method may vary in morphology and size according to the concentration of the second emulsifier of the second aqueous phase.
  • the emulsifier is preferably used in a concentration ranging from 0.25% to 2% (w/v). If the concentration of the emulsifier is less than 0.25%>, the solution being under emulsification becomes unstable, and aggregates of particles are generated. On the other hand, as the concentration of the emulsifier increases within the range, the obtained particles have a smaller particle size and a uniform size distribution.
  • the emulsifier is most preferably used in a concentration of about 2%.
  • the particles prepared by the above method may vary in morphology and size according to the ratio of the magnetic material to the biodegradable synthetic polymer.
  • the magnetic material is preferably used in an amount 0.2 to 0.8 times as much as the biodegradable synthetic polymer.
  • the prepared magnetic nanoparticles are slightly increased in size but greatly improved in encapsulation efficiency into the biodegradable synthetic polymer.
  • the magnetic material is used in an amount identical to or higher than the biodegradable synthetic material, magnetic nanoparticles are formed in a low yield and has a larger particle size. Therefore, in the above method of preparing magnetic nanoparticles according to the present invention, the magnetic material is most preferably used in an amount 0.8 times as much as the biodegradable synthetic polymer.
  • the present inventors found that, in case of using hydrophilic drugs, they are encapsulated with high efficiency by employing the multiple-emulsification technique.
  • the multiple- emulsification technique is preferably employed.
  • the present inventors found that the magnetic nanoparticles prepared by the multiple-emulsification technique have a relatively slow drug release rate.
  • the composition including the magnetic nanoparticles according to the present invention may be administered through various routes known in the art, and may be formulated into various pharmaceutical forms depending on their administration routes.
  • the administration of the present composition may be carried out, for example, intravascularly, intralymphatically, parenterally, subcutaneously, intramuscularly, intranasally, intraperitoneally, interstitially, orally, intratumorly, and others.
  • One preferred route of administration is intravascularly.
  • the present composition is typically injected intravenously, but may be injected intraarterially as well.
  • the present composition may be also injected interstitially or into any body cavity.
  • the present invention provides a targeted drug delivery system capable of specifically concentrating magnetic nanoparticles encapsulating a magnetic material and a drug with a biodegradable synthetic polymer to a target site using a magnetic field.
  • the magnetic nanoparticles are locally concentrated at a target site (that is, a disease site) by application of an external or internal magnetic field to the target site for a predetermined time after administration of the present composition to a subject.
  • the application time of the magnetic field indicates the time consumed for concentrating into a target site 70% of the dosage, preferably 80% of the dosage, and most preferably 90% or higher of the dosage, wherein the time may vary depending on several factors, such as the distance between the administration region, the target site and the administration route.
  • the strength of the magnetic field used for concentrating the magnetic nanoparticles of the present invention to a target site preferably ranges from about 0.2 to 0.3 Tesla.
  • the magnetic nanoparticles concentrated at a target site in this way are able to provide therapeutic efficacy by releasing a drug loaded ' therein simultaneously with degradation by the action of an enzyme in serum.
  • Magnetite Fe 3 O 4
  • the PVA had an average molecular weight of 15,000-20,000 Da and a saponification degree of 88%.
  • Ammonium thiocyanate, hydroperoxide, potassium phosphate (KH 2 PO ) and dibasic potassium phosphate (K 2 HPO 4 ) were purchased from Duksan Pure Chemical Co. (Korea).
  • the prepared particles were found to have defects on their surface and be not uniform in size and mo ⁇ hology.
  • Magnetic nanoparticles were prepared with various amounts of magnetite, ranging from 0.1 to 0.5 g, according to the same method as in Example II except that the ratio of the organic phase to the aqueous phase was 1 :2.
  • the mo ⁇ hology, average size and size distribution of the prepared magnetic nanoparticles were evaluated by using SEM, TEM and DLS. The results are given in FIGS.2, 3 and 4, respectively.
  • the prepared magnetic nanoparticles based on the emulsification-diffiision technique were sleek, well individualized and had a uniform size distribution.
  • the magnetic nanoparticles prepared based on the emulsification-diffiision technique were found to have an average diameter which increased along with the amount of magnetite ranging from 0.1 to 0.5 g, that is, ranging from 150 to 170 nm,
  • the W/O emulsion was added to 20 ml of a second aqueous phase containing PVA in an amount ranging from 0.05 to 0.4 mg, and the resulting solution was emulsified for 1 min using a 15% power probe sonicator, thus giving a W/O/W double emulsion.
  • the W/O/W emulsion was diluted with 100 ml D.D.I water and maintained under gentle agitation. Magnetic nanoparticles thus formed were isolated, as follows.
  • the resulting solution was filtered with a 0.45- ⁇ m membrane. After serum replacement, centrifugation was carried out at 10,000 ⁇ m for 30 min three times. The resulting product was dried in a vacuum oven at 35°C.
  • the magnetic nanoparticles was found to decrease in size and be uniformly individualized as the concentration of the emulsifier in the second aqueous phase increased.
  • concentration of the emulsifier in the second aqueous phase was less than 0.25% (a)
  • aggregates of particles were generated.
  • the use of about 2% emulsifier concentration in the second aqueous phase (b) resulted in production of proper magnetic nanoparticles.
  • Magnetic nanoparticles were prepared with various amounts of magnetite, ranging from 0.1 to 0.5 g, according to the same method as in Example IV except that the concentration of PVA in the second aqueous phase was 2%.
  • the mo ⁇ hology, average size and size distribution of the prepared magnetic nanoparticles were evaluated by using SEM and DLS. The results are given in FIGS. 6 and 7, respectively.
  • the magnetic nanoparticles prepared based on the multiple-emulsification technique had wider size distribution range and were more aggregated than those prepared based on the emulsification-diffiision technique.
  • the magnetic nanoparticles prepared based on the multiple-emulsification technique were found to have an average diameter which increased along with the amount of magnetite ranging from 0.1 to 0.5 g, that is ranging from 350 to 370 nm.
  • the practical concentration of magnetite loaded into the magnetic nanoparticles was determined by measuring absorbance at 480 nm.
  • An HCl/H O 2 (2:3, v/v) solution was added to the magnetic nanoparticles. After oxidization of Fe + to Fe 3 * was allowed to take place, 1 % ammonium thiocyanate was added to the resulting solution. Then, assay for formed thiocyanate complexes was performed by measuring absorbance at 480 nm.
  • Table 1 emulsification-diffiision technique
  • Table 2 multiple-emulsification technique shows the encapsulation efficiency of magnetite into the magnetic nanoparticles which was calculated using the measured absorbance values, and the size of the magnetic nanoparticles below.
  • Both magnetic nanoparticle preparations were found to have an increased particle size along with the amount of magnetite, but this increase in size occurred in small scales.
  • the encapsulation efficiency for magnetite the magnetic nanoparticles were found to vary according to the preparation methods and the amount of magnetite.
  • the maximum encapsulation efficiency for magnetite was found to be respectively about 7.84% for the magnetic nanoparticles prepared based on the emulsification-diffiision technique and about 15.8% for the magnetic nanoparticles prepared based on the multiple-emulsification technique.
  • magnetite loading the amount of magnetite encapsulated into 1 mg of the magnetic nanoparticles
  • encapsulation efficiency ratio of a magnetite loading to a theoretical loading
  • a magnetization curve for magnetite at room temperature did not display any magnetic hysteresis. This result has a correlation with the typical supe ⁇ aramagnetic property of 100 nm magnetite nanoparticles.
  • a magnetic field of 10 kOe or higher basically saturated magnetic powder at room temperature, and a saturation magnetization of 48 emu/g was identified at a magnetic field of 6 kOe.
  • the magnetic nanoparticles were measured to have maximum magnetization of about 10.2 emu/g at a magnetic field of 6 kOe.
  • the curve did not exhibit any hysteresis, remanence and coercitivity.
  • the magnetic nanoparticles were found to have the smaller saturation magnetization than the magnetite with the huge saturation magnetization, but have the paramagnetic property.
  • Drug release from the magnetic nanopaticles were evaluated in a phosphate-buffered aqueous release medium at 37 ⁇ 0.5°C.
  • the dried magnetic nanoparticles were placed into a flask containing 30 ml of the aqueous release medium, and incubated in a shaking incubator (SI-900, J.O. Tech., Korea) at 150 ⁇ m.
  • 3 ml from the aqueous release medium was collected at intervals of 24 hrs, while the medium was supplemented with 3 ml D.D.I. water.
  • the amount of the released drug was monitored by measuring absorbance at 267.8 nm for gemcitabin and at 270.6 nm for cisplatin using an UV spectrometer (UV16A, Shimadzu, Japan) with a standard calibration curve. The results are given in FIGS. 10 and 11.
  • FIGS. 10 and 11 there was a large difference in drug release profiles of the magnetic nanoparticles according to their preparation methods.
  • the magnetic nanoparticles were prepared based on the emulsification-diffiision technique, the magnetic nanoparticles displayed a very rapid drug release rate. It results from the location of the drug which is placed near the surface of the nanoparticles.
  • the preparation of the magnetic nanoparticles based on the multiple- emulsification technique resulted in a very slow drug release. This slow drug release is believed to result from that the drug is located in the polymeric matrix and released by degradation of the polymeric matrix.
  • Encapsulation efficiencies of drugs into the magnetic nanoparticles according to the present invention were investigated by the drug release test of the Example Vm. In this test, total amounts of gemcitabin and cisplatin released from the magnetic nanoparticles for 30 days were measured. The results are given in Table 3, below.
  • the prepared magnetic nanoparticles based on the emulsification- diffiision technique had maximum encapsulation efficiencies of about 18.6% for gemcitabin and about 52.2% for cisplatin. Also, when prepared based on the multiple-emulsification technique, the magnetic nanoparticles had maximum encapsulation efficiencies of about 71.4% for gemcitabin and about 30.4% for cisplatin. Cisplastin (hydrophobic) and gemcitabin (hydrophilic) were found to be encapsulated into the magnetic nanoparticles with high efficiencies by the emulsification-diffiision technique and the multiple-emulsification technique, respectively.
  • HP AC human pancreatic cancer cell line, ATCC No. CRL-2119
  • HP AC human pancreatic cancer cell line, ATCC No. CRL-2119
  • HPAC cells were grown in DMEM:F- 12, which is a mixture of DMEM and Ham's F-12 nutrient medium(l:l) containing 1.2 g/liter NaHCO 3 and 15 mM HEPES.
  • the culture medium was supplemented with 5% fetal bovine serum and 1% antibiotics.
  • Cells were cultured at 37°C in a humidified atmosphere of 5% CO 2 - enriched air, and the medium was replaced every three days.
  • Tumor mass was formed along with the costal ridge of each mouse, while covering most of both flaks of the mice. All mice were maintained under identical conditions.
  • the applied magnetic field strength was determined based on the in vitro study resulted in that a magnetic field of 0.25 Tesla is most effective in targeting the magnetic nanoparticles to the tumor mass.
  • the magnetic nanoparticles according to the present invention were found to be locally concentrated at the magnetic field-applied region.
  • Tumor size was two-dimensionally measured in length and longest width by using calipers. Tumor volume was calculated according to Equation 1 , below, and the results are given as a graph in FIG. 16. Mean values were calculated for each of the test and control groups and compared with each other by an unpaired t test with Welch's correction.
  • the present inventors performed two comparisons for tumor size.
  • the left tumor (applied with the magnetic field) and the right tumor (not applied with the magnetic field) were compared with each other in each mouse of the test group.
  • the second comparison was carried out between the tumor of the control group and the left tumor of the test group.
  • the present inventors found that the magnetic field-applied tumor was more decreased in size than tiie tumor not applied with the magnetic field.
  • the second comparison there was a larger decrease in size of the magnetic field-applied tumor than that of the tumor of the control group.
  • the difference observed in the first comparison was found to be greater than that in the second comparison.
  • the concentration of the magnetic nanoparticles was found to increase in a sequence of the right tumor of the test group, the right and left tumors of the control group and the left tumor of the test group.
  • the anti-tumor effect of the magnetic nanoparticles was found to increase with their increased concentration at the tumor site.
  • the test group, the left tumor was growth- inhibited and degenerated.
  • the composition including the magnetic nanoparticles according to the present invention is capable of substantially concentrating drugs only at disease sites and deeply penetrating into tissues of the disease sites the drugs, thereby minimizing side effects of the drugs and enhancing therapeutic efficacy of the drugs at the disease sites.

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Abstract

Cette invention concerne une composition comprenant des nanoparticules magnétiques, chacune de ces nanoparticules magnétiques enveloppant un matériau magnétique et un médicament à l'aide d'un polymère synthétique biodégradable. Cette invention concerne également un procédé de préparation de ces nanoparticules magnétiques ainsi qu'un système d'administration de médicament ciblé capable de concentrer les nanoparticules magnétiques sur un site cible à l'aide d'un champ magnétique.
PCT/KR2004/001024 2003-04-30 2004-04-30 Composition comprenant une nanoparticule magnetique enveloppant un materiau magnetique et un medicament a l'aide d'un polymere synthetique biodegradable WO2004096190A1 (fr)

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KR1020030027675A KR100612734B1 (ko) 2003-04-30 2003-04-30 자기성 물질 및 치료제를 생분해성 고분자로 캡슐화한자기성 나노입자를 함유하는 조성물

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WO2007105171A2 (fr) * 2006-03-13 2007-09-20 Nannovation Biotech Aps. Elimination de cellules choisies
US20070280418A1 (en) * 2006-06-05 2007-12-06 Sirius Medicine, Llc Coupled Carriers for Enhancing Therapy
WO2008029599A1 (fr) * 2006-08-31 2008-03-13 Canon Kabushiki Kaisha Particules magnétiques et leur procédé de production
EP1996508A1 (fr) * 2006-02-27 2008-12-03 Industry-Academic Cooperation Foundation, Yonsei University Nanoparticules magnétiques ou d'oxyde métallique hydrosolubles couvertes de ligands, et procédé de préparation et utilisation desdites nanoparticules
ES2376941A1 (es) * 2011-11-18 2012-03-21 Universidad De Granada Nanopartículas mixtas de liberación controlada de principios activos.
CN103221334A (zh) * 2010-08-30 2013-07-24 韩华石油化学株式会社 氧化铁纳米胶囊、其制备方法和使用其的mri造影剂
US9345768B2 (en) 2005-04-12 2016-05-24 Magforce Ag Nanoparticle/active ingredient conjugates
WO2019006440A1 (fr) * 2017-06-30 2019-01-03 Otomagnetics, Inc. Nanoparticules magnétiques pour l'administration ciblée

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KR100862973B1 (ko) * 2007-06-28 2008-10-13 연세대학교 산학협력단 표적부위자기약물전달과 조영제를 위한 양이온성 자성나노복합체
KR101125231B1 (ko) * 2009-09-29 2012-03-21 서울대학교산학협력단 초상자성 산화철 나노입자를 포함하는 색전제 및 그 제조 방법
KR101642939B1 (ko) 2010-08-31 2016-07-26 한화케미칼 주식회사 산화철 나노캡슐, 이의 제조방법 및 이를 포함하는 자기공명영상진단 조영제
KR101409296B1 (ko) 2012-09-07 2014-06-24 서울대학교산학협력단 자성 나노 입자의 선택적 활성화 방법 및 선택적 활성화되는 자성 나노 입자
WO2017188725A1 (fr) * 2016-04-29 2017-11-02 서울대학교산학협력단 Dispositif de thermothérapie
KR102048201B1 (ko) * 2017-12-29 2019-11-25 한남대학교 산학협력단 천연색소를 포함하는 나노캡슐 및 이의 제조방법
KR102220657B1 (ko) * 2019-03-12 2021-02-26 재단법인대구경북과학기술원 온열 치료와 약물 방출을 위한 생분해성 마이크로로봇

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US9345768B2 (en) 2005-04-12 2016-05-24 Magforce Ag Nanoparticle/active ingredient conjugates
EP1996508A4 (fr) * 2006-02-27 2010-12-22 Ind Academic Coop Nanoparticules magnétiques ou d'oxyde métallique hydrosolubles couvertes de ligands, et procédé de préparation et utilisation desdites nanoparticules
EP1996508A1 (fr) * 2006-02-27 2008-12-03 Industry-Academic Cooperation Foundation, Yonsei University Nanoparticules magnétiques ou d'oxyde métallique hydrosolubles couvertes de ligands, et procédé de préparation et utilisation desdites nanoparticules
JP2009529004A (ja) * 2006-02-27 2009-08-13 インダストリー−アカデミック コーポレーション ファウンデーション，ヨンセイ ユニバーシティ 相転移リガンドでコートされた水溶性磁性または水溶性金属酸化物ナノ粒子とその製造方法及び用途
WO2007105171A3 (fr) * 2006-03-13 2007-12-13 Nannovation Biotech Aps Elimination de cellules choisies
WO2007105171A2 (fr) * 2006-03-13 2007-09-20 Nannovation Biotech Aps. Elimination de cellules choisies
US20070280418A1 (en) * 2006-06-05 2007-12-06 Sirius Medicine, Llc Coupled Carriers for Enhancing Therapy
WO2008029599A1 (fr) * 2006-08-31 2008-03-13 Canon Kabushiki Kaisha Particules magnétiques et leur procédé de production
JP2008056827A (ja) * 2006-08-31 2008-03-13 Canon Inc 磁性粒子及びその製造方法
CN103221334A (zh) * 2010-08-30 2013-07-24 韩华石油化学株式会社 氧化铁纳米胶囊、其制备方法和使用其的mri造影剂
ES2376941A1 (es) * 2011-11-18 2012-03-21 Universidad De Granada Nanopartículas mixtas de liberación controlada de principios activos.
WO2013072545A1 (fr) * 2011-11-18 2013-05-23 Universidad De Granada Nanoparticules mixtes de libération contrôlée de principes actifs
WO2019006440A1 (fr) * 2017-06-30 2019-01-03 Otomagnetics, Inc. Nanoparticules magnétiques pour l'administration ciblée
JP2020527545A (ja) * 2017-06-30 2020-09-10 オトマグネティクス，インコーポレイテッド 標的化送達のための磁性ナノ粒子

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