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WO2009074274A1 - Nanoparticules polymères solides fonctionnalisées contenant des épothilones - Google Patents

Nanoparticules polymères solides fonctionnalisées contenant des épothilones Download PDF

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Publication number
WO2009074274A1
WO2009074274A1 PCT/EP2008/010371 EP2008010371W WO2009074274A1 WO 2009074274 A1 WO2009074274 A1 WO 2009074274A1 EP 2008010371 W EP2008010371 W EP 2008010371W WO 2009074274 A1 WO2009074274 A1 WO 2009074274A1
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Prior art keywords
dione
polymer
dihydroxy
prop
alkyl
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English (en)
Inventor
Katrin Claudia Fischer
Sascha General
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Bayer Pharma AG
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Bayer Schering Pharma AG
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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
    • A61K9/513—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
    • A61K31/00—Medicinal preparations containing organic active ingredients
    • A61K31/33—Heterocyclic compounds
    • A61K31/395—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins
    • A61K31/41—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having five-membered rings with two or more ring hetero atoms, at least one of which being nitrogen, e.g. tetrazole
    • A61K31/425—Thiazoles
    • A61K31/427—Thiazoles not condensed and containing further heterocyclic rings
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • 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
    • A61K49/0019—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
    • A61K49/0034—Indocyanine green, i.e. ICG, cardiogreen
    • 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
    • A61K49/0039—Coumarin 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
    • A61K49/0054—Macromolecular compounds, i.e. oligomers, polymers, dendrimers
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K49/00—Preparations for testing in vivo
    • A61K49/001—Preparation for luminescence or biological staining
    • A61K49/0063—Preparation for luminescence or biological staining characterised by a special physical or galenical form, e.g. emulsions, microspheres
    • A61K49/0069—Preparation for luminescence or biological staining characterised by a special physical or galenical form, e.g. emulsions, microspheres the agent being in a particular physical galenical form
    • A61K49/0089—Particulate, powder, adsorbate, bead, sphere
    • A61K49/0091—Microparticle, microcapsule, microbubble, microsphere, microbead, i.e. having a size or diameter higher or equal to 1 micrometer
    • A61K49/0093—Nanoparticle, nanocapsule, nanobubble, nanosphere, nanobead, i.e. having a size or diameter smaller than 1 micrometer, e.g. polymeric nanoparticle
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P29/00—Non-central analgesic, antipyretic or antiinflammatory agents, e.g. antirheumatic agents; Non-steroidal antiinflammatory drugs [NSAID]
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P35/00—Antineoplastic agents

Definitions

  • the present invention describes cationic surface potential polymer nanoparticles in which neutral, hydrophobic and hydrophilic pharmaceutically active substances can be included.
  • the hydrophilic and thus water-soluble substances are entrapped by ionic complexation with a charged polymer in the core of the particle by co-precipitation.
  • pharmaceutically active substances include both therapeutics, in particular epothilones, as well as diagnostics.
  • the cationic particle surface provides stable, electrostatic surface modification with partially oppositely charged compounds that may contain target-specific ligands to enhance passive and active targeting.
  • Distribution in the organism can, for. B. for the diagnosis as well as for the therapy of various tumor diseases can be used advantageously.
  • Nanoparticular systems which are used both for detection and for
  • the associated therapeutic monitoring will enable a faster detection of therapy resistance in the future and significantly improve the healing success of the patient through the timely use of alternative therapies
  • a substance class used very successfully in tumor therapy is the group of cytostatic drugs. All rapidly dividing cells of the body, including tumor cells, are damaged by these substances. However, this not only leads to a death of the tumor cells, but often other vital organs and tissues such as the bone marrow, mucous membranes or heart vessels are affected. The associated undesirable toxicity is often the dose limiting factor of therapy [Silacci D., Neri M., Modern Biopharmaceuticals: Design, Development and Optimization, Volume 3, Part V, Wiley-VCH, Weinheim, 2005; 1271-1299].
  • the cardiotoxicity of the substance can be significantly reduced. By reducing the dose-limiting cardiotoxicity, in turn, a higher therapeutic efficiency can be achieved. Due to the demonstrable clinical benefit, the liposome-encapsulated doxorubicin was successfully approved under the name Doxil ® or Cealyx for tumor therapy.
  • Epothilones are a new class of antitumor compounds that cause apoptosis.
  • Various publications and reports of study results eg IDrugs, 2002, 5 (10): 949-954) have proven their effectiveness against cancer.
  • the dose for their administration is also known from study reports or from other publications, eg for epothilones A and B from WO 99/43320.
  • the therapeutic effect or the side-effect profile of this new substance class can be positively influenced by using special distribution mechanisms. First and foremost, the enhanced permeation and retention effect (EPR effect for short) is held responsible.
  • tumor tissue is characterized by its fast growth and various messenger substances usually by a fenestr Understand "holey" tissue structure as well as a lack of lymphatic drainage
  • holey tissue structure As well as a lack of lymphatic drainage
  • this area is also referred to as nanosize window [Hobbs SK, Monsky WL et al., Proc Natl Acad., USA, 1998; 95: 4607-4612] [Brigger I., Dubemet C. et al., Adv. Drug DeNv.
  • the nanoparticles must circulate in the blood stream for a sufficient period of time.
  • This requires particle sizes between about 10 nm and 380 nm and suitable particle surfaces.
  • pegylated particle surfaces can prevent the body's own proteins from identifying the particles as foreign and rapid elimination via the organs of the reticulo-endothelial system (short RES) [Otsuka H. et al., Adv. Drug DeNv. Rev., 2003; 55 (3): 403-419].
  • active ligands on the particle surface eg -A-
  • tissue-specific enrichment can be optimized again [Nobs L. et al., Pharm. Sei., 2004; 93: 1980-1992] [Yokoyama M., J. Artif. Organs, 2005; 8: 77-84].
  • the cell membrane For a recording of the active ingredients in the cell, another physiological barrier, the cell membrane, must be overcome.
  • the active substance is introduced into the cell via endocytosis with the aid of nanoparticles, it is possible to circumvent ejecting transporters and to prevent multi-drug resistance (MDR) [Bharadwaj V., J. Biomed. Nanotechnol., 2005; 1: 235-258] [Huwyler J. et al., J. Drug Target., 2002; 10 (1): 73-79].
  • MDR multi-drug resistance
  • the release properties of the active substance from the nanoparticle can additionally be controlled by targeted selection of the polymer.
  • a nanoparticulate formulation can thus minimize the frequency of application and lead to a reduction of the therapeutically necessary dose. Furthermore, unwanted plasma peak levels can be avoided by encapsulation in nanoparticles and a delayed release can be achieved.
  • tissue or cell specific ligands e.g. B. antibodies
  • controllable drug release by targeted polymer selection
  • a nanoparticulate system which already fulfills all the described advantages, has not yet been developed according to the current state of knowledge.
  • the variety of nanoparticulate carrier systems described in the literature also makes it clear that there is currently no optimal nano-formulation for all problems.
  • the overall structure of the particles, matrix-forming substances, and especially their surface are of crucial importance for in vivo behavior [Choi S.W., Kim W.S., Kim J.H., Journal of Dispersion Science and Technology, 2003; 24 (3 & 4): 475-487].
  • the physicochemical properties of various drugs, especially classes of drugs differ greatly. Accordingly, there remains a need in the development of colloidal drug carrier systems with improved properties.
  • Optical imaging techniques such as sonography, X-ray diagnostics, cross-sectional imaging (CT, MRI) and nuclear medicine (PET, SPECT) are available for in vivo detection.
  • CT cross-sectional imaging
  • PET nuclear medicine
  • SPECT nuclear medicine
  • Another and relatively new method is Optical Imaging, whose detection principle is based on usage is based on near-infrared fluorescence. It is a non-invasive procedure that works without ionizing radiation and is very inexpensive and inexpensive compared to methods such as MRI.
  • the NIR dyes such as indocyanine green developed for such an application are very soluble in water, which makes it difficult to efficiently encapsulate them in a hydrophobic polymer matrix.
  • the reason for this is the rapid change of the hydrophilic substance into the aqueous phase, for example during production by means of nanoprecipitation.
  • hydrophilic substances in nanoparticles only a few technologies are available that have different disadvantages.
  • the amphiphilic nature of liposomes or polymerosomes, for example allows the inclusion of hydrophilic substances in the aqueous interior of the particles, whereas hydrophobic compounds can be incorporated in the membrane. Due to the localization in the core or in the shell of the particles, the loading is very limited and thus usually insufficient.
  • a further disadvantage is that, above all, hydrophilic substances in an aqueous environment are rapidly washed out of such systems.
  • An alternative encapsulation of water-soluble substances in Polyelektrolytkomplexe is limited, since dyes such as indocyanine green (ICG) are small molecules with only a few charged groups, which are insufficient charges for electrostatic complexation available.
  • ICG indocyanine green
  • polyelectrolyte complexes in aqueous solution are very dynamic systems, as a result of which they usually have inadequate colloidal stability in plasma [Thünemann AF et al., Adv. Polym. Sei., 2004; 166: 113-171].
  • both water-soluble dyes for diagnosis and therapeutic substances which are usually poorly water-soluble due to their hydrophobic properties, should be able to be encapsulated effectively and with sufficient stability to wash out.
  • a technological challenge remains to ensure adequate particle stability on the one hand and specific enrichment in the target tissue on the other hand through the use of suitable surfaces.
  • the whereabouts at the place of enrichment (target tissue) is again u. a. depending on how well the particles are absorbed into the tissue and the cell.
  • polymers or substances carrying amino groups are known to possess endo-osmotic activity, ie they promote intracellular activity Release of the particles from the endolysosomes by damage to the endolysosome membrane [DeDuve C. et al., Biochem. Pharmacol., 1974; 23: 2495-2531]. If the particles remain within the cell in the endolysosomes, a degradation of the particle matrix and the substances contained therein takes place via cell-specific enzymes. The endolysosomal release of the encapsulated drugs is therefore essential for the therapeutic effect.
  • nanoparticulate systems with cationic surface properties should be prepared without the potential toxicologically harmful properties hindering an in vivo application.
  • the particle surface must be inconspicuous to the body's own defense mechanisms (Opsonine, RES), allowing only a sufficiently long circulation time, which is a prerequisite for a corresponding accumulation of particles from the blood stream in the target tissue.
  • the nanoparticulate systems should continue to support uptake into the target cell and endolysosomal release.
  • Another difficulty in making nanoparticulate systems is to apply suitable substances or target-recognizing structures to the particle surface.
  • the surface of the particles is modified by means of covalent coupling reactions.
  • Prerequisite for this are functional groups on the polymer backbone or on the particle surface, which can be irreversibly linked by appropriate chemical coupling reactions with the target-recognizing molecule [Nobs L. et al., J.Pharm. Sei., 2004; 93: 1980-1992].
  • the stability of colloidal dispersions is often greatly reduced by the reagents or under the reaction conditions, the chemical implementation is usually complicated and problematic [Koo OM et al., Nanomedicine, 2005; 1 (3): 193-212] [Choi SW et al., J. Dispersion Sei.
  • An object of the invention was therefore to provide an improved pharmaceutical formulation in which both hydrophilic and hydrophobic drugs can be encapsulated.
  • a flexible and sufficiently stable surface modification should allow optimal accumulation in the diseased tissue.
  • such a colloidal system must also be efficiently incorporated into the target tissue as well as into the individual cells where endolysosomal release can occur.
  • they should be practicable manufacturing methods to enable production in a timely and cost-effective manner.
  • the invention relates to polymer nanoparticles having a cationic surface potential containing a cationic polymer and a sparingly water-soluble polymer, characterized in that said polymer nanoparticles encapsulate diagnostic and therapeutic agents, in particular epothilones, together or separately, or contain only epothilones.
  • the cationically functionalized particle surface can be electrostatically stably and flexibly surface-modified with a partially oppositely charged compound.
  • An object of the described invention is therefore the use of nanoparticles according to the invention for the detection of diseases (diagnosis), for the treatment of diseases (therapy) as well as for the monitoring of a therapy.
  • Another aspect of the invention is a kit consisting of separately prepared nanoparticulate systems (a) and (b) thereof
  • a composition comprising (a) a diagnostic agent encapsulated in a particle and (b) a diagnostic agent encapsulated in a particle
  • a further aspect of the invention is a kit as described above, wherein the constituents (a) and (b) are in the solid state, for example as lyophilisate, and if appropriate additionally an agent (c) which is suitable for use in the nanoparticulate systems ( a) and (b) optionally optionally separately or together to disperse.
  • the invention includes the use of the nanoparticulate systems for the preparation of a suitable drug form using pharmaceutically acceptable excipients, which are necessary for the respective dosage form.
  • the drug form developed in the context of the invention can be applied to humans or animals via various routes of administration. Preferably, it is administered iv. Necessary application systems known to the person skilled in the art are likewise part of the invention described here.
  • the composition of the nanoparticles includes a poorly water-soluble polymer, which is preferably a biodegradable polymer or else a mixture of various biodegradable polymers.
  • a biodegradable polymer can be described via individual monomer units which form said polymer by means of polymerization or other processes.
  • the polymer can be defined by its name.
  • the poorly water-soluble polymer is derived from the group of natural and / or synthetic polymers or homo- and / or
  • Methacrylates the lactides, for example, the L-lactides or DL-lactides, the glycolides, the caprolactones such as ⁇ -caprolactones and others from.
  • said polymer or part of the polymer is selected from the group of polycyanoacrylates and polyalkylcyanoacrylates (PACA), such as, for example, polybutylcyanoacrylate (PBCA), polyesters, for example poly (DL-lactides), poly (L-lactides), polyglycolides, polydioxanones, Polyoxazolines, poly (glycolide-co-trimethylene carbonates), polylactide-co-glycolides (PLGA) such as poly (L-lactide-co-glycolide) or poly (DL-lactide-co-glycolide), poly (L-lactide -co-DL-lactides), poly (glycolide-co-trimethylene), poly (carbonates-co-dioxanones), alginic acid, hyaluronic acid, polysialic acid, acidic cellulose derivatives, acidic starch derivatives,
  • PDA polybutylcyanoacrylate
  • polyesters for example
  • Polysaccharides such as dextrans, alginates, cyclodextrins, hyaluronic acid, chitosans, acidic polyamino acids, polymeric proteins such as collagen, gelatin or albumin,
  • Polyamides such as poly (aspartic acid), poly (glutamic acid), poly (lminocarbonate) (poly (carbonate) derived from tyrosine, poly ( ⁇ -hydroxybutyrate),
  • Polyanhydrides such as, for example, polysebacidic acid (poly (SA)), poly (adipic acid), poly (CPP-SA), poly (CPH), poly (CPM) aromatic polyanhydrides, polyorthoesters, polycaprolactones such as poly- ⁇ - or ⁇ -caprolactones,
  • Polyphosphoric acid such as polyphosphates, polyphosphonates, polyphosphazenes, poly (amide-enamines), azopolymers, polyurethanes, dendrimers, Pseudopolymaminoklaren as well as all mixtures and co-polymers of the same compounds.
  • the sparingly water-soluble polymer is selected from the group of the following polymers:
  • Polyacrylates, polylactides and Polygylcolide, or their co-polymers are examples of polyacrylates, polylactides and Polygylcolide, or their co-polymers.
  • the sparingly water-soluble polymer is selected from the group of polyalkyl cyanoacrylates (PACA).
  • PDA polyalkyl cyanoacrylates
  • polyalkylcyanoacrylates shows the structure indicated (formula I), wherein the stated radical R is preferably linear and branched alkyl groups having 1 to 16 carbon atoms, a cyclohexyl, benzyl or a phenyl group.
  • the sparingly water-soluble polymer is a polybutyl cyanoacrylate (PBCA) (formula II).
  • the poorly water-soluble polymer forms the larger proportion of the polymer matrix of the particles.
  • nanoparticles with a cationically charged surface potential are produced by the incorporation of compounds with amino groups, in particular a cationic polymer, into a poorly water-soluble, solid polymer matrix.
  • the cationic polymer is derived from the group of natural and / or synthetic polymers or from homopolymers and copolymers of corresponding monomers.
  • Suitable cationic polymers in the context of this invention are polymers having free primary, secondary or tertiary amino groups which can form salts with any low molecular weight acid, the salts being soluble in aqueous-organic solvents. Also suitable are polymers or their salts which carry quaternary ammonium groups and are soluble in organic solvents.
  • the following groups of cationic polymers, polycations and polyamine compounds or polymers of homo- and co-polymers of corresponding monomers are particularly suitable: modified natural cationic polymers, cationic proteins, synthetic cationic polymers, aminoalkanes of different chain length, modified cationic dextrans, cationic polysaccharides , cationic starch or Cellulose derivatives, chitosans, guar derivatives, cationic cyanoacrylates, methacrylates and methacrylamides, and such monomers and comonomers which can be used to form corresponding suitable compounds and the corresponding salts which can form with suitable inorganic or low molecular weight organic acids.
  • Hydroxyethylcellulose trimethylamine, polyallylamines, protamine chloride, polyallylamine hydrosalts, polyamines, polyvinylbenzyltrimethylammonium salts, polydiallyldimethylammonium salts, polyimidazoline, polyvinylamine and polyvinylpyridine, polyethyleneimine (PEI), putrescine (butane-1, 4-diamine), spermidine (N- (3-aminopropyl) butane-1, 4 -diamine), spermine (N, N'-bis (3-aminopropyl) butan-1, 4-diamine) dimethylaminoethyl acrylate, poly-N, N-dimethylaminoethyl methacrylate P (DMEAMA), dimethylaminopropylacrylamide, dimethylaminopropylmethacrylamide, dimethylaminostyrene, vinylpyridine and Methyldiallylamine, poly-DADMAC, poly
  • polymers are a particular aspect of the invention: polyamines, in particular diethylaminoethyl-modified dextrans, polylysine, protamine sulfate, protamine chloride, polyallylamines and polyallylamine hydrosalts,
  • Polydiallyldimethylammonium salts polyvinylbenzyltrimethylammonium salts, polyimidazoline, polyvinylamine and polyvinylpyridine, polyethyleneimine (PEI), poly-DADMAC, guar, or deacetylated chitin and the corresponding salts which may form with suitable inorganic or low molecular weight organic acids.
  • the following monomers are suitable: hydroxymethylcellulosetrimethylamine, hydroxyethylcellulosetrimethylamine, putrescine (butane-1, 4-diamine), spermidine (N- (3-aminopropyl) butane-1,4-diamine), spermine (N, N'-bis (3 - aminopropyl) butane-1, 4-diamine), dimethylaminoethyl acrylate,
  • Suitable acids for salt formation are, for. B.: Hydrochloric acid, sulfuric acid, but especially acetic acid, glycolic acid or lactic acid.
  • the amino group-bearing compound in particular a cationic polymer, in an organic solvent which is infinitely miscible with water, preferably acetone, methanol, ethanol, propanol, dimethyl sulfoxide (DMSO), or in a mixture of these solvents with water be solved.
  • an organic solvent which is infinitely miscible with water, preferably acetone, methanol, ethanol, propanol, dimethyl sulfoxide (DMSO), or in a mixture of these solvents with water
  • the polymer nanoparticles contain, as the amino group bearing compound, a cationically modified polyacrylate (poly-N, N-dimethylaminoethyl methacrylate, P (DMAEMA)) (Formula 3).
  • a cationically modified polyacrylate poly-N, N-dimethylaminoethyl methacrylate, P (DMAEMA)
  • the biodegradable, cationically modified polyacrylate eg P (DMAEMA), P (DMAPMAM) is encapsulated in the polymer matrix, in particular the PBCA polymer matrix, by nanoprecipitation.
  • the surface of the resulting nanoparticles has a positive (cationic) surface potential (zeta potential) due to the amino groups of the cationic polymer.
  • the cationic particle surface ensures good cell uptake and enables flexible electrostatic surface modification with partially anionically charged compounds.
  • the polymer nanoparticles contain as cationic polymer a modified polyacrylate poly (dimethylaminopropylmethacrylamide (P (DMAPMAM)).
  • DMAPMAM dimethylaminopropylmethacrylamide
  • the polymer nanoparticles contain as cationic polymer polyethyleneimine (PEI) of various molecular weights, in particular 1, 8 kDa, 10 kDa, 70 kDa and 750 kDa, (formula 4).
  • PEI polyethyleneimine
  • PEI is a polycation commonly used in the field of non-viral gene therapy for DNA polyplexes (PEK) and accordingly much studied [Remy J.-S. et al., Adv. Drug Deliv. Rev., 1998; 30 (1-3): 85-95].
  • the particle shell consists of PEI polymer chains, which produce a cationic surface potential.
  • diagnostic as well as therapeutic substances can be included in the polymer matrix by means of nanoprecipitation.
  • the following substance classes for different molecular imaging methods can be used as diagnostic substances for encapsulation, in particular contrast agents or tracers for the following method for molecular imaging (molecular imaging): optical imaging, e.g. DOT (diffuse optical imaging), US (ultrasound imaging), OPT (optical projection tomography), near infrared fluorescence imaging, fluorescence protein imaging and BLI (bioluminescence imaging) and magnetic resonance imaging (MRI, MRI) and X-ray imaging (X-ray).
  • optical imaging e.g. DOT (diffuse optical imaging), US (ultrasound imaging), OPT (optical projection tomography), near infrared fluorescence imaging, fluorescence protein imaging and BLI (bioluminescence imaging) and magnetic resonance imaging (MRI, MRI) and X-ray imaging (X-ray).
  • DOT direct optical imaging
  • US ultrasound imaging
  • OPT optical projection tomography
  • near infrared fluorescence imaging fluorescence protein imaging
  • BLI bioluminescence
  • nanoparticulate systems according to the invention which contain exclusively a diagnostic active substance are not covered by the patent claims.
  • the diagnostic agent is a dye, in particular selected from the following group: fluorescein, fluorescein isothiocyanate, carboxyfluorescein or calcein, tetrabromfluoresceins or eosines, tertaiodefluorescein or erythrosines, difluorofluorescein, such as Oregon GreenTM 488, Oregon GreenTM 500 or Oregon GreenTM 514, Carboxyrhodol (Rhodol Green TM) dyes (U.S. 5,227,487, U.S. 5,442,045), carboxyrodamine dyes (Rhodamine Green TM Dyes) (U.S.
  • the diagnostic substance is a fluorescent dye.
  • the diagnostic agent is a fluorescent near-infrared (NIR) dye.
  • NIR dyes preferably used for optical imaging, absorb and emit light in the NIR range between 650 nm and 1000 nm.
  • the preferred dyes belong to the class of polymethine dyes and are selected from the following groups: carbocyanines such as diethyloxacarbocyanine (DOC), diethyloxadicarbocyanine (DODC ), Diethyloxatricarbocyanine (DOTC), indodi or indotricarbocyanines, tricarbocyanines, merocyanines, oxonol dyes (WO 96/17628), rhodamine dyes, phenoxazine or phenothiazine dyes,
  • carbocyanines such as diethyloxacarbocyanine (DOC), diethyloxadicarbocyanine (DODC ), Diethyloxatricarbocyanine (DOTC), in
  • Tetrapyrrole dyes especially benzoporphyrins, chorine and phthalocyanines.
  • Suitable inorganic cations or counterions for these dyes are, for example, the lithium ion, the potassium ion, the hydrogen ion and in particular the sodium ion.
  • Suitable cations of organic bases include those of primary, secondary or tertiary amines, such as ethanolamine, diethanolamine, morpholine, glucamine, N, N-dimethylglucamine, and especially N-methylglucamine and polyethyleneimine.
  • Suitable cations of amino acids are, for example, those of lysine, arginine and ornithine and the amides of otherwise acidic or neutral amino acids.
  • the dyes can be used as bases or as salts of these.
  • the diagnostic substance is a carbocyanine dye.
  • the general structure of the carbocyanines is described as follows: (Formula VI)
  • R 30 represents a hydrogen atom, a hydroxy group, a carboxy group, an alkoxy group having 1 to 4 carbon atoms or a chlorine atom
  • R 31 represents a hydrogen atom or an alkyl group having 1 to 4 carbon atoms
  • R 20 to R 29 , R 32 and R 33 independently of one another represent a hydrogen atom, a hydroxy group, a carboxy, a sulfonic acid radical or a carboxyalkyl, alkoxycarbonyl or alkoxyoxoalkyl radical having up to 10 C atoms or a sulfoalkyl radical up to 4 carbon atoms, or for a non-specifically binding macromolecule, or for a radical of the general formula (VII)
  • R 20 to R 29 is a non-specifically binding macromolecule or the general formula VII wherein o and s are 0 or independently of one another are an integer from 1 to 6, q and v independently of one another are 0 or 1, R 34 represents a hydrogen atom or a methyl radical,
  • R 35 is an alkyl radical having 3 to 6 C atoms which has 2 to n-1 hydroxyl groups, where n is the number of C atoms, or an alkyl radical having 1 to 6 C atoms which is substituted by 1 to 3 carboxy groups,
  • carbocyanines In the case of carbocyanines, further reference is made to the applications DE 4445065 and DE 69911034, the content of which should also be the subject of this application.
  • the use of the carbocyanines mentioned there for the nanoparticles of the present invention is a particular aspect in the context of the present invention.
  • anionic, readily water-soluble substances such as certain carbocyanines can be stably entrapped in the hydrophobic polymer matrix of the described nanoparticles.
  • an anionic water-soluble substance is encapsulated by ion complexation and co-precipitation with a cationic polymer in a sparingly water-soluble polymer matrix by nanoprecipitation, resulting in particles of defined size.
  • NIR-active fluorescent dye By incorporating an NIR-active fluorescent dye into the polymer matrix of the particles, they are non-invasively detectable by fluorescence in the tissue by means of optical imaging. There is thus the possibility in vivo of detecting the distribution or accumulation of fluorescently labeled nanoparticles.
  • the carbocyanine dye is the readily water-soluble anionic tetrasulfocyanine (TSC) (Formula IX).
  • the carbocyanine dye is IDCC (indodicarbocyanine) (Formula IX).
  • the carbocyanine dye is ICG (indocyanine green) (Formula XI).
  • the encapsulated pharmaceutically active substance is an epothilone.
  • the epothilone is defined by the general formula (XII)
  • R 1a , R 1b are each independently of one another hydrogen, C 1 -C 10 -alkyl, aryl, aralkyl, or together a group - (CH 2 ) m - wherein m is 2 to 5;
  • R 2a R 2b independently of one another are hydrogen, C 1 -C 10 -alkyl, C 2 -C 10 -alkenyl, C 2 -C 10 -alkynyl, aryl, aralkyl, or together form a group - (CH 2 ) n - where n is 2 to 5 .
  • R J is hydrogen, C 1 -C 0 alkyl, aryl, aralkyl
  • R, 4 ⁇ a, R 4b independently of one another are hydrogen, C 1 -C 10 -alkyl, aryl, aralkyl, or together form a group - (CH 2 ) P - where p is 2 to 5;
  • R b is hydrogen, C 1 -C 10 -alkyl, aryl, aralkyl, CO 2 H, CO 2 alkyl, CH 2 OH, CH 2 O-C 1 -C 5 -alkyl, CH 2 OAcyl, CN, CH 2 NH 2 , CH 2 N ((C r C 5 alkyl), acyl) i, 2, or CH 2 Hal, Chai 3;
  • R 6 , R 7 are independently hydrogen, or together one
  • DE can not be CH 2 -O;
  • R 8 is hydrogen, C 1 -C 10 alkyl, aryl, aralkyl, halogen, CN;
  • R 9 , R 10 independently of one another are hydrogen, C 1 -C 20 -alkyl, aryl,
  • Aralkyl or together with the methylene carbon atom, a 5- to 7-membered carbocyclic ring;
  • R 11 is hydrogen or a protective group PG Z ;
  • R 12 is hydrogen, or C 1 -C 10 alkyl
  • PG Z CrC 2O alkyl a C 4 -C 7 cycloalkyl group which may contain one or more oxygen atoms in the ring, aryl, aralkyl, C 1 -C 20 acyl, aroyl, C 1 -C 20 alkylsulfonyl, arylsulfonyl, tri (C 1 -C 20 alkyl) silyl, di ( C 1 -C 20 alkyl) arylsilyl, (C 1 -C 20 alkyl) diarylsilyl, or tri (aralkyl) silyl;
  • the epothilone is defined by the formula (XII) wherein R 1a , R 1b independently of one another are hydrogen, C 1 -C 4 -alkyl, or together form the group - ⁇ CH 2 ) m - where m is 2 to 5;
  • R 2a , R 2b independently of one another are hydrogen, C 1 -C 8 -alkyl, or together form the group - ⁇ CH 2 ) m - where m is 2 to 5, or C 2 -C 6 -alkenyl, or C 2 -C 6 -alkynyl ;
  • R 3 is hydrogen
  • R 48 , R 4b independently of one another are hydrogen, C 1 -C 4 -alkyl,
  • R 5 is hydrogen, C 1 -C 4 -alkyl.
  • R 6 , R 7 are both hydrogen, or together another bond, or together an epoxide function
  • DE is the group H 2 C-CH 2 , or
  • X is the group CR 9 R 10 ;
  • R 8 is hydrogen, C 1 -C 4 -alkyl, halogen,
  • R 9, R 10 are both independently hydrogen, C r C 4 alkyl,
  • R 12 is hydrogen, or C 1 -C 4 -alkyl; encapsulated as a single stereoisomer or as a mixture of different stereoisomers and / or as pharmaceutically acceptable salts.
  • R 1a , R 1b are both independently of one another hydrogen, C 1 -C 2 -alkyl, or together form a group - (CH 2 ) m - where m is 2 to 5,
  • R 2a , R 2b are each independently of one another hydrogen, C 1 -C 5 alkyl, or together the group a - (CH 2 ) n - wherein n is 2 to 5, or C 2 -C 6 alkenyl, or C 2 -C 6 alkynyl; R 3 is hydrogen,
  • R 48 , R 4b are each, independently of one another, hydrogen, C 1 -C 2 -alkyl, R 5 is hydrogen or methyl or trifluoromethyl,
  • R 6 , R 7 together form another bond, or together an epoxide function
  • DE is the group H 2 C-CH 2 .
  • X is the group CR 9 R 10 ;
  • R 8 is hydrogen, methyl, chlorine, fluorine,
  • R 9 , R 10 are both independently of one another hydrogen, C 1 -C 4 -alkyl,
  • R 12 is hydrogen or C 1 -C 4 -alkyl; encapsulated as a single stereoisomer or as a mixture of different stereoisomers and / or as a pharmaceutically acceptable salt.
  • epothilones of general formula (XII) are encapsulated, which are listed in the following list: (4S, 7R, 8S, 9S, 13E / Z, 16S) -4,8-dihydroxy-16- (2-methyl -benzoxazol-5-yl) -1-oxo-5,5,9,1-tetramethyl-phi-en-i-ylJ-cyclohexadec-IS-ene-1-dione;
  • the use of the nanoparticles of the invention containing an epothilone for the treatment of tumors diseases associated with inflammatory reactions is an aspect of the invention.
  • Another aspect of the invention is a method of treating tumors or diseases associated with inflammatory reactions in which an effective amount of the active ingredient contained in a nanoparticulate system according to the invention is administered to humans or animals.
  • the diagnostic agent and the epothilone are incorporated together in the particle. Accordingly, the use of these nanoparticles for the treatment of tumor diseases or diseases associated with inflammatory reactions as well as for the simultaneous diagnosis and / or monitoring of the course of therapy is an aspect of the invention. In particular, therapy and monitoring is an aspect of the present invention.
  • a further aspect of the invention is therefore a method for the treatment, diagnosis and / or monitoring of tumor diseases or diseases associated with inflammatory reactions in which an effective amount of the active substance contained in a nanoparticulate system according to the invention is administered to humans or animals.
  • a method of treatment and monitoring of therapy is one aspect of the invention.
  • the diagnostic agent and the epothilone are present in separate identical nanoparticulate systems, which in particular have the same structure.
  • the invention thus also relates to a kit consisting of the particles according to the invention which together contain a diagnostic agent and an epothilone and a kit which contains (a) the particles according to the invention comprising a diagnostic agent and (b) the particles according to the invention comprising an epothilone separately, and Use of the kit for therapy and diagnosis / monitoring.
  • the polymer nanoparticles according to the claims are precipitation aggregates which are produced by nanoprecipitation.
  • the following production methods are available for this purpose: Direct precipitation (precipitation) in a test tube by introducing the dissolved polymer-substance mixture into a surfactant-containing aqueous solution which is mixed by means of a magnetic stirrer.
  • the organic solvent is abruptly withdrawn from the matrix polymer and the substances dissolved with it, when the polymer-containing organic solution is added to a significantly larger volume of an aqueous solution.
  • compounds dissolved in the polymer phase with amino groups are co-encapsulated in the sparingly soluble polymer during the precipitation.
  • the condition for this is the unlimited miscibility of the organic solvent (particularly suitable, for example, acetone, ethanol) with water and the insolubility of the matrix polymer in the aqueous phase.
  • the diagnostic agent is negatively charged and trapped in the particle as an ion pair with the cationic polymer.
  • the surface of the polymer nanoparticles is electrostatically modified.
  • the electrostatic modification of the cationic nanoparticle surface is an outstanding advantage of the present invention.
  • the particle surface can be modified with a suitable substance without a chemical coupling reaction.
  • the prerequisite for this is that the modifying agent has partial charges which are opposite to the particle surface charge.
  • electrostatic surface modification by charge titration allows a simple, flexible and versatile modification of the particle surface.
  • the enrichment (active and passive targeting) of nanoparticles from the bloodstream into the target tissue requires that the particles circulate in the blood stream for a sufficient period of time.
  • the circulation time in the body can be adjusted individually, in particular by using polyethylene oxides or polyethylene glycols (see Example 5).
  • electrostatic surface modification described here can be carried out quickly and without problems directly before use. This is done by simply mixing appropriate amounts of the nanoparticle dispersion with the modifying agent. Thus, it is additionally possible to prepare and store the core particles separately from the surface-modifying agent. This is for a particularly advantageous for the colloidal long-term stability. On the other hand, extremely labile surface-modifying substances such as peptides or antibodies can be stored under suitable conditions until use.
  • the separation of core particles and modifying agent further allows a surface modification according to the individual requirements of the patient.
  • the surface modification on a modular principle offers maximum flexibility for diagnosis, therapy and monitoring, whereby the uncomplicated implementation of the modification is carried out directly by the user.
  • a preferred construction of the surface-modifying agent for cationically functionalized polymer nanoparticles, in particular the described PBCA nanoparticles, is shown in formula 5.
  • the partially anionically charged moiety fulfills the function of an anchor on the positively charged particle surface through electrostatic interactions.
  • the neutral, aligned to the surrounding aqueous medium moiety consists of polyethylene glycol and / or polyethylene oxide units (PEG units) of different lengths.
  • PEG chains having a molecular weight of from 100 to 30,000 daltons and more preferably from 3,000 to 5,000 daltons. This moiety may alternatively be made of other suitable structures, such as.
  • HES hydroxyethyl starch
  • the radical R is preferably hydrogen or a methyl unit.
  • the anionic anchor may be, for example, a polymer of 5 to 50 units of glutamic acid (Glu) or aspartic acid (Asp) or the salts of the acids. It may also be a mixed polymer of the named subunits. Furthermore, uncharged subunits such. B. neutral amino acids in the anionic block regularly or irregularly incorporated. Generally suitable as a negatively charged moiety (anchor) compounds or polymeric structures with groups such as acetate, carbonate, citrate, succinate, nitrate, carboxylate, phosphate, sulfonate or sulfate groups, as well as salts and free acids of these Groups.
  • the anionic anchor may preferably be a polyamino acid.
  • the anionic anchor consists of a polymer of up to 20 units of glutamic acid (Glu) or aspartic acid (Asp) or its salts, or their copolymers, which may optionally also contain neutral amino acids.
  • the surface is modified with Glu (10) -b-PEG (110), Glu (10) -b-PEG (114) or Asp (15) -b-PEG (114).
  • the surface is modified with Glu (10) -b-PEG (110) or Asp (15) -b-PEG (114).
  • the surface of the polymer nanoparticle is modified with Glu (10) -b-PEG (110) (Formula XIV).
  • the negative moiety (anchor) is the carboxylate groups of the glutamic acid subunits of the block copolymer.
  • Formula XIV Structural Formula of GIu (10) -b-PEG (110);
  • a target recognizing structure is present.
  • This target recognizing structure has at least one negatively charged moiety which is applied to the cationic particle surface by electrostatic interactions.
  • Another particularly preferred structure of the surface-modifying agent for cationically functionalized polymethylene particles, in particular the described PBCA nanoparticles, is shown in formula XV.
  • the anionic anchor may be constructed according to the previously described description of formula XIII.
  • the partially anionically charged moiety fulfills the function of an anchor on the positively charged particle surface through electrostatic interactions.
  • the middle, neutral part of the molecule consists of polyethylene glycol units and / or polyethylene oxide units (PEG units) of different lengths. Preference is given here to PEG chains having a molecular weight of from 100 to 30,000 daltons and more preferably from 3,000 to 5,000 daltons. This moiety may alternatively be made of other suitable structures, such as. As hydroxyethyl starch (HES) and all possible polymeric compounds exist.
  • HES hydroxyethyl starch
  • the ligand X of the surface-modifying agent also called the target-recognizing structure in the following, serves to improve passive and active accumulation mechanisms of the polymanneroparticles.
  • Suitable ligands X as targeting structures may be antibodies, peptides, receptor ligands of ligand mimetics or an aptamer. Structures include amino acids, peptides, CDR (compementary determining regions), antigens, haptens, enzyme substances, enzyme cofactors, biotin, carotenoids, hormones, vitamins, growth factors, lymphokines, carbohydrates, oligosaccharides, lecithins, dextrans, lipids, nucleosides such as DNA or an RNA molecule containing native, modified or artificial nucleosides, Nucleic acids, oligo nucleotides, polysaccharides, B, A, Z HeNx or hairpin structure (hairpin), modified polysaccharides as well as receptor binding substances or fragments thereof. Target-recognizing structures may also be transferrin or folic acid or parts thereof, or all possible combinations of the foregoing.
  • the targeting structure is selected from a list containing an antibody, a protein, a polypeptide, a polysaccharide, a DNA molecule, an RNA molecule, a chemical moiety, a nucleic acid, a lipid, a carbohydrate or combinations of the foregoing.
  • ligands are bound to the nanoparticles via electrostatic interactions, but it is also possible to bind the ligands via covalent bonds to the particle surface. Furthermore, it is possible to incorporate a linker between ligand and nanoparticles.
  • the electrostatic attachment of the target-recognizing structures takes place via charge interactions with at least one negatively charged moiety on the cationic particle surface.
  • a negatively charged moiety are compounds or polymeric structures having groups such as acetate, carbonate, citrate, succinate, nitrate, carboxylate, phosphate, sulfonate or sulfate groups, as well as salts and free acids of these groups ,
  • the size of the nanoparticles is between 1 nm and 800 nm.
  • the size of the nanoparticles is between 5 nm and 800 nm.
  • the size of the nanoparticles is between 1 nm and 500 nm. In a preferred embodiment, the size of the nanoparticles is between 1 nm and 300 nm.
  • the size of the nanoparticles is between 5 nm and 500 nm.
  • the size of the nanoparticles is between 5 nm and 300 nm.
  • the size of the nanoparticles is between 10 nm and 300 nm.
  • the size of the resulting polymer nanoparticles is determined by photon correlation spectroscopy (PCS).
  • the preparation of the polymer nanoparticles is characterized by carrying out the following process steps:
  • the water-insoluble polymer is dissolved in a suitable organic solvent
  • Solvent which is infinitely miscible with water preferably acetone, methanol, ethanol, propanol, isopropanol dimethyl sulfoxide (DMSO), or dissolved in a mixture of these solvents with water.
  • the cationic polymer is dissolved in a suitable solvent which is infinitely miscible with water, preferably acetone, methanol, ethanol, propanol, dimethylsulfoxide (DMSO), or dissolved in a mixture of these solvents with water.
  • the active ingredient (diagnostic agent and / or epothilone) is dissolved in an organic solvent which is infinitely miscible with water, preferably acetone, methanol, ethanol, propanol, dimethylsulfoxide (DMSO), isopropanol or in a mixture of these solvents with water.
  • an organic solvent which is infinitely miscible with water, preferably acetone, methanol, ethanol, propanol, dimethylsulfoxide (DMSO), isopropanol or in a mixture of these solvents with water.
  • DMSO dimethylsulfoxide
  • the organic solvent is then removed either under atmospheric pressure or negative pressure, via lyophilization or heat completely or other suitable methods.
  • the aqueous stable nanoparticle dispersion prepared above is mixed in a suitable proportion with the modifying agent dissolved in water.
  • the determination of the appropriate quantitative ratio is carried out by stepwise titration of the particle dispersion with the modifying agent.
  • the degree of electrostatic surface modification (charge titration) is controlled by determining the zeta potential.
  • the target value of the zeta potential depends on the planned application path. For i.v. Applications would be preferred, for example, a neutral to negative zeta potential, in oral and buccal administration, a neutral or cationic zeta potential is preferred, etc.
  • a purification step is carried out, for example by washing the particles with a suitable solution.
  • Suitable solutions for the washing process are, for example, aqueous surfactant solutions 0.1-2%, but also pure water.
  • lyophilized nanoparticles after removal of the organic solvent and optionally purification or after Surface modification are lyophilized.
  • the lyophilized nanoparticles may then be marketed as a kit and reconstituted and administered for use.
  • kits which encapsulates the particles containing a diagnostic agent and an epothilone or only one epothilone together or separately, optionally as a lyophilisate or as a solution or dispersion.
  • This kit may additionally comprise a suitable means for reconstitution of the lyophilisate, e.g. physiological saline or water for injection / infusion purposes.
  • Another object of the invention is a kit that the non-surface-modified particles, means for the preparation of a solution of the surface-modifying agent.
  • the described nanoparticles can be further processed using suitable pharmaceutical additives to form various dosage forms which are suitable for application to humans or animals.
  • suitable pharmaceutical additives include in particular aqueous dispersions, lyophilisates, solid oral dosage forms such as fast-dissolving tablets, capsules and others.
  • suitable pharmaceutical additives may be: sugar alcohols for lyophilization (eg sorbitol, mannitol), tableting excipients, polyethylene glycols, etc.
  • aqueous nanoparticle dispersion or a further developed pharmaceutical form can be administered orally, parenterally (eg intravenously), subcutaneously, intramuscularly, intraocularly, intrapulmonary, nasally, intraperitoneally, dermally as well as on all other administration routes possible for humans or animals.
  • parenterally eg intravenously
  • subcutaneously intramuscularly, intraocularly, intrapulmonary, nasally, intraperitoneally, dermally as well as on all other administration routes possible for humans or animals.
  • the invention relates to a process for the preparation of a polymer nanoparticle, characterized in that the following process steps are carried out:
  • active agent includes therapeutically and diagnostically-effective compounds, as well as compounds that are active in animals other than humans and plants.
  • Epothilone or epothilone encompasses all naturally occurring epothilones and their derivatives Epothilone derivatives are known, for example, from WO 93/10102, WO 93/10121 and DE 41 38 042 A2, WO 97/19086 and WO 98/25929, WO 99 / 43320, WO 2000/066589, WO 00/49021, WO 00/71521, WO 2001027308, WO 99/02514, WO 2002080846.
  • epothilones suitable for use in the present invention and their preparation are disclosed in DE 19907588, WO 98/25929, WO 99/58534, WO 99/2514, WO 99/67252, WO 99/67253, WO 99/7692, EP 99/4915, WO 00/1333, WO 00/66589, WO 00/49019, WO 00/49020, WO 00/49021, WO 00/71521, WO 00/37473, WO 00/57874, WO 01/92255, WO 01/81342, WO 01/73103, WO 01/64650, WO 01/70716, US 6204388, US 6387927, US 6380394, US 02/52028, US 02/58286, US 02/62030, WO 02/32844, WO 02/30356, WO 02/32844, WO 02/14323, and WO 02/8440. Particularly suitable are the compounds disclosed in WO 00/665
  • epothilones as encompassed by formula II.
  • the term epothilone also includes the possibility of encapsulating in a preparation various epothilone derivatives selected from the disclosed list of claim 21.
  • matrix polymer as used herein describes the polymer which quantitatively constitutes the major proportion of the particle mass, wherein further encapsulated substances (both arbitrary additives and pharmaceutically active substances) may be uniformly and / or non-uniformly embedded.
  • (nano) precipitation describes the formation of a colloidal precipitate by precipitating a sparingly water-soluble polymer when placed in an aqueous phase, with mixing of the solvents taking place Community precipitation of several substances which may be both water-soluble and poorly water-soluble in the context of the invention.
  • this precipitating aggregate consists of a matrix polymer in which further polymeric substances as well as pharmaceutically active substances can be partially or completely embedded in. A uniform or uneven distribution of the coagulum may be present. encapsulated substances in the matrix polymer.
  • anchor describes an ionic moiety of the modifying agent that allows immobilization and thus localization of the modifying agent on the charged particle surface by ionic interactions between oppositely charged compounds.
  • Charge titration describes the process of electrostatic coupling of the armature on the particle surface that can be traced by zeta potential measurement.
  • the charged anchor changes the zeta potential of the particle towards the charge of the anchor.
  • neutral amino acid in the sense of the invention encompasses all amino acids whose molecular charge is balanced in the sum of the individual charges, ie zero, including all natural amino acids R-CH (COOH) (NH 2), whose radical R carries no charge eg alanine, valine, leucine, isoleucine, etc. but also synthetic amino acids that fulfill this condition, such as cyclohexylalanine.
  • surfactants according to the invention are, on the one hand, surface-active substances which lower the interfacial tension between two immiscible phases, which makes it possible to stabilize colloidal dispersions.
  • surfactants can be any type of substances which are capable of sterically and / or electrostatically stabilizing colloidal dispersions.
  • tumors includes the diseases or conditions associated with cell growth, cell division and / or proliferation, such as malignant tumors More specific diseases which fall into this term include, for example, osteoporosis, osteoarthritis, cervical and cervical cancer, and abdominal cancer including all carcinomas in the upper abdomen, colon cancer, glandular cancer eg adenosarcoma, breast, lung (eg SCLC and NSCLC), head and neck carcinoma, malignant melanoma, acute lymphocytic or myelocytic leukemia, prostate carcinoma, bone metastases, brain tumors and brain metastases Solid tumors are a sub-aspect. Preferred among “tumor diseases” are neonatal and cervical cancer, breast, lung, prostate carcinoma, malignant melanoma, bone metastases, brain tumors and brain metastases to understand.
  • targeting is meant a targeting structure: in the case of “passive targeting”, the targeting structure may indirectly assist in enrichment at the target site, for example, by extending the circulation time of the particles in the bloodstream increases the likelihood of enrichment about the fenestrations in the tumor endothelium.
  • active targeting is used when tissue- or cell-specific ligands are used for targeted enrichment Active ligands can be administered directly to active substances (ligand-drug Conjugates) as well as to the surface of colloidal carrier systems.
  • passive targeting is used when the distribution of the active ingredient is due to (nonspecific) physical, biochemical or immunological processes, primarily due to the enhanced permeation and retention effect (EPR effect) this is a passive enrichment mechanism which exploits the structural peculiarities of tumorous or inflamed tissue [Ulbrich K., Subr V., Adv. Drug DeNv. Rev., 2004; 56 (7): 1023-
  • surface potential also referred to as surface charge
  • zeta potential is synonymous with the term “zeta potential”. This zeta potential is determined by the method of laser Doppler anemometry (LDA).
  • the surface potential also referred to as zeta potential, indicates the potential of a migrating particle at the shear plane, i. H. if the majority of the diffuse layer has been sheared off by movement of the particle.
  • the surface potential was determined by the method of laser Doppler anemometry using a "Zetasizer 3000" (Malvern Instruments).
  • the migration speed of the particles in the electric field is determined. Particles with a charged surface migrate in an electric field to the oppositely charged electrode, wherein the
  • Migration rate of the particles depends on the amount of surface charges and the applied field strength.
  • particles traveling in the electric field are irradiated with a laser and the scattered laser light is detected.
  • a frequency shift in the reflected light is measured in comparison to the incident light.
  • Doppler frequency Doppler effect
  • the electrophoretic mobility results from the quotient of the migration speed and the electric field strength.
  • the product of electrophoretic mobility and factor 13 corresponds to the zeta potential, the unit of which is [mV].
  • the software used was PCS V1.41 / PCS V1.51 Rev.
  • the control measurements of the zeta potential were carried out with standard latex particles from Malvern Instruments Ltd. (-50 mV ⁇ 5 mV). The measurements were made under the standard settings of Malvern Instruments Ltd. carried out.
  • the size of the nanoparticles was determined by Dynamic Light Scattering (DLS) using a "Zetasizer 3000" (Malvern Instruments). In addition, images were taken in the scanning electron microscope (SEM), as shown by way of example in FIG. Figure 12 ( Figure 12) also confirms the spherical shape of the nanoparticles.
  • the determination of the particle size by DLS is based on the principle of Photon Correlation Spectroscopy (PCS).
  • PCS Photon Correlation Spectroscopy
  • the particle size is calculated from the slope of the correlation function.
  • the particles should have a spherical shape, which can be checked by SEM images (see above), do not sediment or float.
  • the measurements were carried out with samples in appropriate dilution, at a constant temperature of 25 ° C and a defined viscosity of the solution.
  • the measuring device was calibrated with standard latex particles of different sizes from Malvern Instruments Ltd.
  • the scanning electron micrographs (SEM images) for determining the particle size were prepared using a field emission scanning electron microscope of the type XL-30-SFEG from FEI (Kassel, Germany). In advance, the samples were sputtered in a high vacuum sputter 208 HR from Cressington (Watford, England) with a 5 nm gold-palladium layer.
  • the solubility of a substance indicates whether and to what extent a pure substance can be dissolved in a solvent. It thus describes the property of a substance to mix with the solvent under homogeneous distribution (as atoms, molecules or ions).
  • the solubility of a compound is determined to be the concentration of a saturated solution that is in equilibrium with the undissolved sediment, depending on the temperature (room temperature).
  • a sparingly soluble compound has a solubility ⁇ 0.1 mol / l, a moderately soluble between 0.1-1 mol / l and a slightly soluble compound> 1 mol / l.
  • Sicomet 6000 is used for PBCA production by anionic polymerization of butyl cyanoacrylate (BCA).
  • BCA butyl cyanoacrylate
  • the polymerization process is carried out by slow, permanent dropwise addition of a total of 2.5% [m / v] BCA in a 1% [m / v] Triton X-100 solution in an acidic solution (pH 1, 5 - pH 2.5 , ideal pH 2.2).
  • the pH is previously adjusted by means of a 0.1 N HCl solution.
  • the resulting dispersion is stirred while cooling in an ice bath (about 4 ° C) for 4 hours constantly at 450 U / min. Subsequently, larger agglomerates are separated by filtration through a paper pleated filter.
  • the BCA polymerized to PBCA is precipitated and the resulting filter cake is washed several times with purified water (MilliQ system). After the drying of the PBCA filter residue in a drying oven at 40 0 C for 24 h by GPC a number average molecular weight determined (Mn ⁇ 2000 Da). Polystyrene standards are used.
  • Example 2 Production of Epothilone-loaded PBCA-P (DMAEMA) Nanoparticles by Nanoprecipitation
  • the PBCA used is prepared by Example 1. 10 ml of a 1% Synperonic T707 solution (m / v) are placed in a 20 ml screw-capped magnetic stirrer tube.
  • the PBCA used is prepared by Example 1.
  • Crystalline, unencapsulated epothilone is separated by means of a 1 ⁇ m glass fiber filter (PALL, 25 mm, 1 ⁇ m P / N 4523T).
  • the filtrate is purified using an Amicon ultrafiltration cell 8050 using a 100 kDa polyethersulfone filter membrane.
  • the filtrate is concentrated to about 1/3 of the volume and then the concentrate is brought back to the starting volume by adding a 1% Synperonic T707 solution. It follows a renewed narrowing. This process is repeated twice and the target concentration of the epothilone particle dispersion is adjusted via the final volume of the solution (epothilone content 0.1-2 mg / ml).
  • PBCA-P DMAEMA nanoparticles (with ICG, DODC, IDCC or Coumarin 6)
  • Dye solution a 3 mg of indocyanine green are pre-dissolved in 300 ⁇ l of purified water in an ultrasonic bath and then treated with 700 ⁇ l of acetone.
  • Dye Solution b, c, d The dyes DODC, IDCC and coumarin 6 are used in a 0.02% acetone solution [m / v].
  • the mixed dye-polymer mixture is taken up in a 2.5 ml Eppendorf pipette and in 10 ml of an intensely stirred 1% [m / v] Synperonic T707 solution pipetted.
  • the nanoparticle dispersion is stirred for 2 h at 600 rpm (standard magnetic stirrer) and for a further 16 h at 100 rpm for complete evaporation of the solvent.
  • the workup is carried out by centrifugation in Eppendorf Caps.
  • the mixed dye-polymer mixture is taken up with a 2.5 ml Eppendorf pipette and pipetted into 10 ml of an intensely stirred 1% Triton X-100 solution.
  • the nanoparticle dispersion is stirred for 2 h at 600 rpm (standard magnetic stirrer) and for a further 16 h at 100 rpm for complete evaporation of the solvent.
  • the workup is carried out by centrifugation in Eppendorf Caps.
  • Example 6 Influence of Nanoprecipitation by Changing the Polymer Content in the Surfactant Phase
  • FIG. 4 shows that the particle size of the PBCA-P (DMAEMA) nanoparticles can be controlled during production by varying the polymer concentration.
  • the stabilization of the PBCA-P (DMAEMA) nanoparticles prepared according to Example 2 (but without epothilone) is carried out with the Synperonic T707 surfactant.
  • the volume of the organic polymer solution injected into the surfactant phase is kept constant and only the polymer concentration is changed accordingly. All further
  • Example 7 Electrostatic surface modification of epothilone-loaded PBCA-PDMAEMA nanoparticles with Glu (10) -b-PEG (110)
  • the epothilone-PBCA-PDMAEMA nanoparticles used here are prepared according to Example 2.
  • the aqueous, stable nanoparticle dispersion is mixed in a suitable quantitative ratio with the modifying agent (Glu (10) -b-PEG (110) / Glu (10) -b-PEG (114) dissolved in water).
  • the determination of the appropriate quantitative ratio is carried out by stepwise titration of the particle dispersion with the modifying agent.
  • the degree of electrostatic surface modification (charge titration) is controlled by determining the zeta potential.
  • Fig. 3a Shown in Fig. 3a) is the change of the zeta potential from (+) 25 mV to about (-) 30 mV, or Fig. 3b) from +35 mV to -10 mV, by stepwise addition of the modifying agent (Glu (10) -b-PEG (110)) or Glu (10) -b-PEG (114) for particle dispersion (charge titration).
  • the modifying agent Glu (10) -b-PEG (110)
  • Example 8 SEM images of epothilone-loaded PBCA-P (DMAEMA) nanoparticles
  • the epothilone-loaded PBCA-P (DMAEMA) are prepared according to Example 2.
  • FIG. 5 shows an SEM micrograph of epothilone-loaded PBCA-P (DMAEMA) nanoparticles.
  • FCS fetal calf medium
  • Additives penicillin / streptamycin
  • the cells are passaged regularly and sowed for experimental purposes 24 hours before the start of the investigations.
  • the cells are in 96-well plates of the company
  • FCS-containing medium is aspirated and replaced with 50 .mu.l of serum-free medium.
  • CMXRos dye previously diluted in medium, from Molecular Probes Europe BV, Leiden (NL) (0.25 ⁇ l / ml) is used. Incubation with 50 ⁇ l of the dye solution takes place in the incubator for 15 min (37 ° C., 5% CO 2 ). Then the dye solution is aspirated and the cells with 2-3 times
  • PBS washed.
  • the fixation of the cells is carried out with 100 ul of 1, 37% formaldehyde for 10 min at room temperature. After aspirating the fixing solution, the cells are washed 2-3 times with PBS. Nuclear staining takes place on the already fixed cells with a maximum of 33342.
  • 100 ⁇ l of the dye solution diluted in PBS (2 ⁇ g / ml) are incubated for 10 min at room temperature. After removing the dye solution, the cells are washed with 100 ⁇ l PBS 2-3 times.
  • the fixed plates are stored in the refrigerator until the fluorescence microscopic examination with 200 .mu.l PBS / Well protected from light in the refrigerator at 8 ° C.
  • Example 9 The nanoparticles used in Example 9 are prepared according to Example 5. Unmodified or after electrostatic surface modification with folic acid or Glu (10) -b-PEG (110), the particles have the properties listed in Table 1.
  • Example 11 Cell uptake behavior Glu (10) -b-PEG (110) modified PBCA P (DMAEMA) nanoparticle loaded with coumarin 6 (fluorescent dye for in vitro detection)
  • the brightly fluorescent dots which are endosomes or endolysosomes, are evidence of efficient uptake of the nanoparticles into the cell by endocytosis ( Figure 7).
  • the scale of the magnification proves that in this photograph individual particles can not be visible due to their size of less than 200 nm.
  • a large number of particles within these vesicles cause strong, punctate fluorescence contrast in the cytoplasm.
  • Cell uptake of PBCA-P (DMAEMA) nanoparticles surface-modified with Glu (10) -b-PEG (110) is shown schematically in FIG.
  • Example 12 Enrichment of the GIu (10) -b-PEG (110) Modified PBCA P (DMAEMA) Nanoparticles in the Nucleus
  • GIu (10) -b-PEG (110) Modified PBCA P (DMAEMA) Nanoparticles in the Nucleus On the basis of the recording of the central cell plane with the aid of the confocal laser scanning microscope (FIG. 9), it can be shown that partial accumulation of the particles takes place in the cell nucleus.
  • Charge titration is used to modify the cationic surface of the PBCA-P (DMAEMA) nanoparticles with the block copolymer Glu (10) -b-PEG (110) (see FIG. 14).
  • the surface charge measured as zeta potential will correspondingly increase from approx. +3 ohms above the neutral point until it reaches its neutral point of the dissociation equilibrium at about -30 mV.
  • the surface-modified PBCA [P (DMAEMA) -ICG] particles show no change in the zeta potential over the investigated period of 7 days after titration. In connection with the unchanged particle size and the constant low PI, a good particle stability can be proven.
  • FIG. 15 shows the UV-Vis absorption spectra of an aqueous ICG solution as well as the ICG nanoparticle dispersion (washed and unwashed).
  • Indocyanine green is a near-infrared fluorescent dye whose absorption and emission spectrum is in the wavelength range between 650-900 nm.
  • DMAEMA cationic polyacrylate P
  • the animals used are supplied by Taconic M & B. These are female albino nude mice of the type NMRI nude. The adult animals have a weight of 22-24 g after about 8 weeks.
  • Five female nude mice are inoculated with 2x10 6 cells of a F9 teratoma in the right posterior flank. The cells are obtained from the company ATCC / LGC Promochem GmbH. It is mouse-derived embryonic cells of a testinal teratocarcinoma, which is used as a tumor model for cancer research purposes in mice. After 18 days, four of the five mice had tumors with an average size of about 0.5-1 cm in diameter.
  • the animals are permanently anesthetized with a rompun-ketavet injection in a dose of 100 ⁇ l / 10 g of animal for the first hour of the experiment.
  • the solution for injection consists of a 1: 1 mixture of a 1:10 dilution Rompun or 1: 5 dilution Ketavet with physiological saline.
  • 200 ⁇ l of the nanoparticle dispersion are injected iv into the tail vein.
  • the subsequent anesthesia is carried out with Rompun- Ketavet via the lungs as an inhalation anesthetic to the circulation of the animals only minimal weight.
  • the animals are examined by fluorescence optics.
  • GIu (10) -b-PEG (110) -modified PBCA [P (DMAEMA) -ICG] nanoparticles after intravenous administration (tail vein) are capable of passive enrichment mechanisms (EPR effect) in tumor tissue.
  • Examination of the tumors ex vivo shows a clear enhancement of the fluorescence contrast in treated compared to untreated tumor tissue (compare Fig. 18 b with a and c with a).
  • Multiple, time-shifted detection of fluorescence in one and the same animal is possible after 24 h and 48 h (FIG. 17). Consequently, the particles can circulate in vivo for a sufficiently long time and accumulate accordingly in the tumor.
  • the electrostatically pegylated surface is thus stably connected to the particle surface.
  • the device used for the animal experiment was set up by the company LMTB (Berlin, Germany). As individual components were used:
  • Excitation filter 1xLCLS-750 nm-F; 1x740 nm interference filter (bandpass)
  • Emission filter 1 x bk-802.5-22-C1; 1 xbk-801 -15-C1
  • Fig. 1 Short-term stability of PBCA-PDMAEMA nanoparticles with epothilone (not surface-modified).
  • Fig. 2 Short-term stability PBCA-PDMAEMA nanoparticles with epothilone surface-modified with Glu (10) -b-PEG (110) a) Hydrodynamic particle diameter d hyd and polydispersity index PI / # EpoPDI 9Ak2konz PEG-Glu b) Hydrodynamic particle diameter d h y d and Zeta potential
  • Fig. 3 Titration curve (zeta potential) in the surface modification of epothilone-loaded PBCA-PDMAEMA nanoparticles. Shown in this figure is the change of the
  • Fig. 4 control of the particle diameter by changing the
  • Fig. 1 it is shown that the particle size of the PBCA-P (DMAEMA) nanoparticles can be controlled during production by varying the polymer concentration.
  • FIG. 5 The figure shows an SEM image of epothilone-loaded PBCA-P (DMAEMA) nanoparticles.
  • Cell uptake a) Comparison of cell uptake behavior after surface modification; Row 1: unmodified particles; Row 2: Nanoparticles with folic acid; Row 3: Nanoparticles with Glu (10) -b-
  • Fig. 7 Nanoparticle uptake in HeLa cells; Fluorescence of
  • the figure shows the cell uptake behavior of GIu (10) -b-PEG (HO) modified PBCA P (DMAEMA) nanoparticles in HeLa.
  • PEG-NP pegylated coumarin-containing PBCA-P (DMAEMA) nanoparticles
  • NP coumarin-loaded PBCA-P (DMAEMA) nanoparticles
  • CP clathrin-coated pits
  • ES endosomes
  • LS lysosomes
  • ELS endolysosomes
  • ZK cell nucleus
  • H + H + ATPase
  • PEG-Glu free Glu (10) -b-PEG (110)
  • the plot shows the accumulation of the GIu (10) -b-PEG (110) modified PBCA-P (DMAEMA) nanoparticles in the nucleus. This is possible by loading with the fluorescence-active dye coumarin 6.
  • Fig. 10 Lower particle uptake with incubation of lower particle concentration: 0.21 mg / ml; Fluorescence of the NP as grayscale image; The figure shows fluorescent HeLa cells after a particle concentration of 0.21 mg / ml was incubated.
  • GIu (10) -b-PEG (110) surface-modified PBCA-P (DMAEMA) particles were used for this.
  • Fig. 11 Increased particle uptake with incubation of higher particle concentration: 0.85 mg / ml; Fluorescence of the NP as grayscale image;
  • the figure shows more strongly fluorescent HeLa cells after a higher particle concentration of 0.85 mg / ml was incubated.
  • GIu (10) -b-PEG (110) surface-modified PBCA-P (DMAEMA) particles were used for this purpose.
  • Fig. 12 SEM image of PBCA [P (DMAEMA) ICG] nanoparticles
  • FIG. 13 particle diameter d hyd of the PBCA [P (DMAEMA) ICG] nanoparticles, surface-modified with Glu (10) -b-PEG (110); The particle size of the surface-modified PBCA [P (DMAEMA) -ICG] nanoparticles used for the animal experiment is shown over a period of 7 days
  • Figure 14 Zeta potential of the untitrated (washed / unwashed) and titrated PBCA [P (DMAEMA) ICG] nanoparticles; The figure shows the zeta potential measured
  • PBCA-P (DMAEMA) nanoparticles modified with the block copolymer Glu (10) -b-PEG (110). This was correspondingly titrated from about + 3OmV beyond the neutral point until reaching the dissociation equilibrium at about -3OmV.
  • Fig. 15 UV-Vis absorption spectra: a) aqueous ICG solution, b) PBCA [P (DMAEMA) -ICG] -NP unwashed; c) PBCA- [P (DMAEMA) -ICG] nanoparticles washed;
  • Fig. 16 Emission spectrum of the PBCA [P (DMAEMA) ICG] nanoparticles and an aqueous ICG solution; The figure represents the corresponding emission spectra of the aqueous ICG solution compared to the nanoparticle dispersion.
  • Fig. 17 Detection of NIR fluorescence in vivo; The images show the NIR fluorescence in a time frame of 24 and 48 h after substance injection (a) ventrally 24 h, b) 24 h laterally, c) 48 h laterally, d) ventrally empty).
  • Fig. 18 NIR fluorescence contrast of the tumor tissue ex vivo 48 h after treatment
  • the figure shows NIR fluorescence contrasts a) an untreated tumor without NIR fluorescence contrast, b) a large, treated tumor and c) a median, treated tumor ex vivo 48 h after treatment.

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Abstract

L'invention concerne des nanoparticules polymères à potentiel de surface cationique, pouvant contenir des substances pharmaceutiquement actives tant hydrophobes qu'hydrophiles. Les substances hydrophiles et donc hydrosolubles sont incluses dans le coer de la particule par coprécipitation, par complexion ionique avec un polymère chargé. Pour l'encapsulation, on peut utiliser comme substances pharmaceutiquement actives tant des produits thérapeutiques que des produits de diagnostic. La surface cationique des particules permet une modification de surface électrostatique stable, avec des composés partiellement à charges opposées qui, afin d'améliorer le ciblage passif et actif, peuvent contenir des ligands spécifiques de la cible.
PCT/EP2008/010371 2007-12-10 2008-12-06 Nanoparticules polymères solides fonctionnalisées contenant des épothilones Ceased WO2009074274A1 (fr)

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Cited By (15)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2012026608A1 (fr) * 2010-08-24 2012-03-01 Canon Kabushiki Kaisha Particule polymère et colorant hydrophile ayant un groupe sulfonate encapsulé dans la particule
JP2013514381A (ja) * 2009-12-15 2013-04-25 バインド バイオサイエンシズ インコーポレイテッド エポチロンを含む治療用ポリマーナノ粒子ならびにそれを製造および使用する方法
EP2404598A4 (fr) * 2009-03-03 2013-10-23 Public Univ Corp Yokohama City Particules de polymère de cyanoacrylate conjuguées à un acide aminé
US8603534B2 (en) 2008-06-16 2013-12-10 Bind Therapeutics, Inc. Drug loaded polymeric nanoparticles and methods of making and using same
US8603535B2 (en) 2009-12-11 2013-12-10 Bind Therapeutics, Inc. Stable formulations for lyophilizing therapeutic particles
US8623417B1 (en) 2008-06-16 2014-01-07 Bind Therapeutics, Inc. Therapeutic polymeric nanoparticles with mTOR inhibitors and methods of making and using same
US8905997B2 (en) 2008-12-12 2014-12-09 Bind Therapeutics, Inc. Therapeutic particles suitable for parenteral administration and methods of making and using same
US8912212B2 (en) 2009-12-15 2014-12-16 Bind Therapeutics, Inc. Therapeutic polymeric nanoparticle compositions with high glass transition temperature or high molecular weight copolymers
US9198874B2 (en) 2008-12-15 2015-12-01 Bind Therapeutics, Inc. Long circulating nanoparticles for sustained release of therapeutic agents
US9351933B2 (en) 2008-06-16 2016-05-31 Bind Therapeutics, Inc. Therapeutic polymeric nanoparticles comprising vinca alkaloids and methods of making and using same
US9877923B2 (en) 2012-09-17 2018-01-30 Pfizer Inc. Process for preparing therapeutic nanoparticles
US9895378B2 (en) 2014-03-14 2018-02-20 Pfizer Inc. Therapeutic nanoparticles comprising a therapeutic agent and methods of making and using the same
EP3566719A1 (fr) 2010-05-18 2019-11-13 Cerulean Pharma Inc. Compositions et procédés pour le traitement de maladies auto-immunes et d'autres maladies
US10758623B2 (en) 2013-12-09 2020-09-01 Durect Corporation Pharmaceutically active agent complexes, polymer complexes, and compositions and methods involving the same
CN115078292A (zh) * 2021-03-10 2022-09-20 北京大学 一种水性超分子粘附材料及其制备方法

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* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2015181346A1 (fr) * 2014-05-30 2015-12-03 AbbVie Deutschland GmbH & Co. KG Nanocapsules de poly(2-cyanoacrylate d'alkyle) hautement chargées en médicament
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CN119768461A (zh) * 2022-08-22 2025-04-04 爱克发-格法特公司 近红外和红光吸收复合树脂颗粒

Citations (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO1990013256A1 (fr) * 1989-05-04 1990-11-15 Wisconsin Alumni Research Foundation Procede de localisation et de traitement de tumeurs a l'aide de nouveaux complexes
WO1999018934A1 (fr) * 1997-10-09 1999-04-22 Vanderbilt University Dispositif d'administration de microparticules ou de nanoparticules de polymeres
WO2007025767A2 (fr) * 2005-09-02 2007-03-08 Schering Ag Complexe nanoparticulaire d'inclusion et de charge pour formulations pharmaceutiques
WO2007141050A2 (fr) * 2006-06-08 2007-12-13 Bayer Schering Pharma Aktiengesellschaft Nanoparticules polymères solides fonctionnalisées pour applications diagnostiques et thérapeutiques

Family Cites Families (44)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US262030A (en) 1882-08-01 Wash-stand
US252028A (en) 1882-01-10 Alonzo s
US258286A (en) 1882-05-23 Fruit-drier
US4774339A (en) 1987-08-10 1988-09-27 Molecular Probes, Inc. Chemically reactive dipyrrometheneboron difluoride dyes
US5366860A (en) 1989-09-29 1994-11-22 Applied Biosystems, Inc. Spectrally resolvable rhodamine dyes for nucleic acid sequence determination
US5433896A (en) 1994-05-20 1995-07-18 Molecular Probes, Inc. Dibenzopyrrometheneboron difluoride dyes
US5227487A (en) 1990-04-16 1993-07-13 Molecular Probes, Inc. Certain tricyclic and pentacyclic-hetero nitrogen rhodol dyes
US5274113A (en) 1991-11-01 1993-12-28 Molecular Probes, Inc. Long wavelength chemically reactive dipyrrometheneboron difluoride dyes and conjugates
US5248782A (en) 1990-12-18 1993-09-28 Molecular Probes, Inc. Long wavelength heteroaryl-substituted dipyrrometheneboron difluoride dyes
US5187288A (en) 1991-05-22 1993-02-16 Molecular Probes, Inc. Ethenyl-substituted dipyrrometheneboron difluoride dyes and their synthesis
DE4138042C2 (de) 1991-11-19 1993-10-14 Biotechnolog Forschung Gmbh Epothilone, deren Herstellungsverfahren sowie diese Verbindungen enthaltende Mittel
FR2684103B1 (fr) 1991-11-22 1994-09-02 Centre Nat Rech Scient Nouveaux composes a structure guanidique et composition pharmaceutique les contenant.
DE4445065A1 (de) 1994-12-07 1996-06-13 Diagnostikforschung Inst Verfahren zur In-vivo-Diagnostik mittels NIR-Strahlung
JP4183099B2 (ja) 1995-11-17 2008-11-19 ゲゼルシャフト・フュア・ビオテヒノロジッシェ・フォルシュング・ミット・ベシュレンクテル・ハフツング（ゲー・ベー・エフ） エポチロンｃおよびｄ、製造法ならびに組成物
US6204388B1 (en) 1996-12-03 2001-03-20 Sloan-Kettering Institute For Cancer Research Synthesis of epothilones, intermediates thereto and analogues thereof
US6441186B1 (en) 1996-12-13 2002-08-27 The Scripps Research Institute Epothilone analogs
US6380394B1 (en) 1996-12-13 2002-04-30 The Scripps Research Institute Epothilone analogs
DE19728914A1 (de) 1997-07-07 1999-01-14 Basf Ag Metallisch gefärbte Polyamide
US6605599B1 (en) 1997-07-08 2003-08-12 Bristol-Myers Squibb Company Epothilone derivatives
WO1999007692A2 (fr) 1997-08-09 1999-02-18 Schering Aktiengesellschaft Nouveaux derives d'epothilone, leur procede de fabrication et leur utilisation pharmaceutique
US6320045B1 (en) 1997-12-04 2001-11-20 Bristol-Myers Squibb Company Process for the reduction of oxiranyl epothilones to olefinic epothilones
FR2775187B1 (fr) 1998-02-25 2003-02-21 Novartis Ag Utilisation de l'epothilone b pour la fabrication d'une preparation pharmaceutique antiproliferative et d'une composition comprenant l'epothilone b comme agent antiproliferatif in vivo
DE19820599A1 (de) 1998-05-08 1999-11-11 Biotechnolog Forschung Gmbh Epothilonderivate, Verfahren zu deren Herstellung und deren Verwendung
AU4775299A (en) 1998-06-22 2000-01-10 Nicolaou, Kyriacos Costa Desmethyl epothilones
SE517641C2 (sv) 1998-07-02 2002-07-02 Sca Hygiene Prod Ab Förfarande för att framställa ett materialskikt med barriärflikar samt ett ytskikt med sådana barriärflikar
JP2000095758A (ja) 1998-09-18 2000-04-04 Schering Ag 近赤外蛍光造影剤および蛍光造影方法
EP1140944B1 (fr) 1998-12-22 2003-08-27 Novartis AG Derives d'epothilone et leur utilisation comme agents antitumoraux
DE19908763A1 (de) 1999-02-18 2000-08-24 Schering Ag Neue Epothilon-Derivate, Verfahren zu deren Herstellung und ihre pharmazeutische Verwendung
DE19908760A1 (de) 1999-02-18 2000-08-24 Schering Ag Neue Epothilon-Derivate, Verfahren zu deren Herstellung und ihre pharmazeutische Verwendung
SK11852001A3 (sk) 1999-02-18 2002-04-04 Schering Aktiengesellschaft Deriváty epotiólonu, spôsoby ich výroby a ich farmaceutické použitie
DE19907588A1 (de) 1999-02-22 2000-08-24 Biotechnolog Forschung Gmbh C-21 Modifizierte Epothilone
IL144590A0 (en) 1999-03-29 2002-05-23 Bristol Myers Squibb Co A process for the preparation of epothilone derivatives
AR023792A1 (es) 1999-04-30 2002-09-04 Bayer Schering Pharma Ag Derivados 6-alquenilo- y 6-alquinilo-epotilona, los procedimientos para prepararlos y su empleo en productos farmaceuticos
CA2425620A1 (fr) 1999-10-08 2001-04-19 The Scripps Research Institute Derives d'epothilone d'alkyle-13 inferieur
EP1259490A2 (fr) 2000-03-01 2002-11-27 Sloan Kettering Institute For Cancer Research Synthese d'epothilones, de leurs produits intermediaires et de leurs analogues
US6518421B1 (en) 2000-03-20 2003-02-11 Bristol-Myers Squibb Company Process for the preparation of epothilone analogs
US6593115B2 (en) 2000-03-24 2003-07-15 Bristol-Myers Squibb Co. Preparation of epothilone intermediates
DE10020517A1 (de) 2000-04-19 2001-10-25 Schering Ag Neue Epothilon-Derivate, Verfahren zu deren Herstellung und ihre pharmazeutische Verwendung
US20020045609A1 (en) 2000-05-26 2002-04-18 Gary Ashley Epothilone derivatives and methods for making and using the same
US20020137152A1 (en) 2000-07-25 2002-09-26 Daniel Santi Fermentation process for epothilones
UA75365C2 (en) 2000-08-16 2006-04-17 Bristol Myers Squibb Co Epothilone analog polymorph modifications, a method for obtaining thereof (variants), a pharmaceutical composition based thereon
CA2425828A1 (fr) 2000-10-13 2002-04-18 The University Of Mississippi Synthese d'epothilones et d'analogues associes
ATE349412T1 (de) 2000-10-16 2007-01-15 R & D Biopharmaceuticals Epothilon-synthesebausteine i: unsymmetrisch substituierte acyloine und acyloinderivate, verfahren zu deren herstellung sowie deren verwendung zur herstellung von epothilonen und epothilonderivaten
WO2002080846A2 (fr) 2001-04-03 2002-10-17 Kosan Biosciences, Inc. Derives d'epothilone : procedes de fabrication et methodes d'utilisation associes

Patent Citations (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO1990013256A1 (fr) * 1989-05-04 1990-11-15 Wisconsin Alumni Research Foundation Procede de localisation et de traitement de tumeurs a l'aide de nouveaux complexes
WO1999018934A1 (fr) * 1997-10-09 1999-04-22 Vanderbilt University Dispositif d'administration de microparticules ou de nanoparticules de polymeres
WO2007025767A2 (fr) * 2005-09-02 2007-03-08 Schering Ag Complexe nanoparticulaire d'inclusion et de charge pour formulations pharmaceutiques
WO2007141050A2 (fr) * 2006-06-08 2007-12-13 Bayer Schering Pharma Aktiengesellschaft Nanoparticules polymères solides fonctionnalisées pour applications diagnostiques et thérapeutiques

Non-Patent Citations (1)

* Cited by examiner, † Cited by third party
Title
YANG S C ET AL: "Formation of positively charged poly(butyl cyanoacrylate) nanoparticles stabilized with chitosan", COLLOID & POLYMER SCIENCE, SPRINGER VERLAG, HEIDELBERG, DE, vol. 278, no. 4, 1 January 2000 (2000-01-01), pages 285 - 292, XP002481607, ISSN: 0303-402X *

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EP2512459A4 (fr) * 2009-12-15 2013-08-07 Nanoparticules polymères thérapeutiques comportant de l'épothilone et leurs procédés de fabrication et d'utilisation
US9835572B2 (en) 2009-12-15 2017-12-05 Pfizer Inc. Therapeutic polymeric nanoparticle compositions with high glass transition temperature or high molecular weight copolymers
EP3566719A1 (fr) 2010-05-18 2019-11-13 Cerulean Pharma Inc. Compositions et procédés pour le traitement de maladies auto-immunes et d'autres maladies
US9610367B2 (en) 2010-08-24 2017-04-04 Canon Kabushiki Kaisha Particle and contrast agent having the particle
WO2012026608A1 (fr) * 2010-08-24 2012-03-01 Canon Kabushiki Kaisha Particule polymère et colorant hydrophile ayant un groupe sulfonate encapsulé dans la particule
US9877923B2 (en) 2012-09-17 2018-01-30 Pfizer Inc. Process for preparing therapeutic nanoparticles
US10758623B2 (en) 2013-12-09 2020-09-01 Durect Corporation Pharmaceutically active agent complexes, polymer complexes, and compositions and methods involving the same
US11529420B2 (en) 2013-12-09 2022-12-20 Durect Corporation Pharmaceutically active agent complexes, polymer complexes, and compositions and methods involving the same
US9895378B2 (en) 2014-03-14 2018-02-20 Pfizer Inc. Therapeutic nanoparticles comprising a therapeutic agent and methods of making and using the same
US10071100B2 (en) 2014-03-14 2018-09-11 Pfizer Inc. Therapeutic nanoparticles comprising a therapeutic agent and methods of making and using the same
CN115078292A (zh) * 2021-03-10 2022-09-20 北京大学 一种水性超分子粘附材料及其制备方法

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TW200932220A (en) 2009-08-01
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PE20091107A1 (es) 2009-08-29
PA8806901A1 (es) 2009-07-23
AR069638A1 (es) 2010-02-10

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