WO2005010481A2 - Preparation et utilisation de glyconanoparticules en or - Google Patents

Preparation et utilisation de glyconanoparticules en or Download PDF

Info

Publication number
WO2005010481A2
WO2005010481A2 PCT/US2004/017607 US2004017607W WO2005010481A2 WO 2005010481 A2 WO2005010481 A2 WO 2005010481A2 US 2004017607 W US2004017607 W US 2004017607W WO 2005010481 A2 WO2005010481 A2 WO 2005010481A2
Authority
WO
WIPO (PCT)
Prior art keywords
ligands
attached
core portion
gold
nanoparticle
Prior art date
Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
Ceased
Application number
PCT/US2004/017607
Other languages
English (en)
Other versions
WO2005010481A3 (fr
Inventor
Jacquelyn Gervay-Hague
Birte Nolting
Current Assignee (The listed assignees may be inaccurate. Google has not performed a legal analysis and makes no representation or warranty as to the accuracy of the list.)
University of California Berkeley
University of California San Diego UCSD
Original Assignee
University of California Berkeley
University of California San Diego UCSD
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Application filed by University of California Berkeley, University of California San Diego UCSD filed Critical University of California Berkeley
Publication of WO2005010481A2 publication Critical patent/WO2005010481A2/fr
Publication of WO2005010481A3 publication Critical patent/WO2005010481A3/fr
Anticipated expiration legal-status Critical
Ceased legal-status Critical Current

Links

Classifications

    • 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
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K33/00—Medicinal preparations containing inorganic active ingredients
    • A61K33/24—Heavy metals; Compounds thereof
    • A61K33/242—Gold; Compounds thereof
    • 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
    • A61K31/70—Carbohydrates; Sugars; Derivatives thereof
    • A61K31/715—Polysaccharides, i.e. having more than five saccharide radicals attached to each other by glycosidic linkages; Derivatives thereof, e.g. ethers, esters
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K33/00—Medicinal preparations containing inorganic active ingredients
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K33/00—Medicinal preparations containing inorganic active ingredients
    • A61K33/24—Heavy metals; Compounds thereof
    • A61K33/243—Platinum; Compounds thereof
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K47/00—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient
    • A61K47/50—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates
    • A61K47/51—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the non-active ingredient being a modifying agent
    • A61K47/54—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the non-active ingredient being a modifying agent the modifying agent being an organic compound
    • A61K47/549—Sugars, nucleosides, nucleotides or nucleic acids
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K47/00—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient
    • A61K47/50—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates
    • A61K47/69—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the conjugate being characterised by physical or galenical forms, e.g. emulsion, particle, inclusion complex, stent or kit
    • A61K47/6921—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the conjugate being characterised by physical or galenical forms, e.g. emulsion, particle, inclusion complex, stent or kit the form being a particulate, a powder, an adsorbate, a bead or a sphere
    • A61K47/6923—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the conjugate being characterised by physical or galenical forms, e.g. emulsion, particle, inclusion complex, stent or kit the form being a particulate, a powder, an adsorbate, a bead or a sphere the form being an inorganic particle, e.g. ceramic particles, silica particles, ferrite or synsorb
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K47/00—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient
    • A61K47/50—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates
    • A61K47/69—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the conjugate being characterised by physical or galenical forms, e.g. emulsion, particle, inclusion complex, stent or kit
    • A61K47/6921—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the conjugate being characterised by physical or galenical forms, e.g. emulsion, particle, inclusion complex, stent or kit the form being a particulate, a powder, an adsorbate, a bead or a sphere
    • A61K47/6927—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the conjugate being characterised by physical or galenical forms, e.g. emulsion, particle, inclusion complex, stent or kit the form being a particulate, a powder, an adsorbate, a bead or a sphere the form being a solid microparticle having no hollow or gas-filled cores
    • A61K47/6929—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the conjugate being characterised by physical or galenical forms, e.g. emulsion, particle, inclusion complex, stent or kit the form being a particulate, a powder, an adsorbate, a bead or a sphere the form being a solid microparticle having no hollow or gas-filled cores the form being a nanoparticle, e.g. an immuno-nanoparticle

Definitions

  • the HIV virus gains entry into host cells through a cascade of events mediated by the viral glycoproteins gpl20 and gp41 (see, (a) Allan, J. S.; Coligan, J. E.; Barin, F.; McLane, M. F.; Sodroski, J. G.; Rosen, C. A.; Haseltine, W. A.; Lee, T. H.; Essex, M. Science 1985, 228, 1091-1094; (b) Barin, F.; McLane, M. F.; Allan, J. S.; Lee, T. H.; Groopman, J. E.; Essex, M. Science 1985, 228, 1094-1096; and (c) McKeating, J.
  • T-lymphocytes and macrophages are the primary targets of HIN infection and the process is initiated by interactions between gpl20, CD4, and the chemokine co-receptor. These interactions lead to conformational changes in gpl20, which subsequently unmask a hydrophobic region of gp41 that is capable of inserting into the host cell membrane.
  • GalCer is a glycosphmgolipid (GSL) expressed on mucosal membrane cells that do not express CD4.
  • GSL glycosphmgolipid
  • McReynolds, et al. have shown that gpl20 (recombinant gpl20 is used in most studies) recognizes several different GSLs in addition to GalCer including naturally occurring GlcCer, and LacCer and even non-natural analogs such as MelCer, and CelCer ( Figure 1, see, McReynolds, K.
  • compositions to facilitate the .study of cell recognition and adhesion processes, that are useful in systems comprising the complete panoply of enzymes present in vivo. Additionally, compositions are needed that can be used in therapeutic applications in which enzymatic degradation contributes to reduced half-life and which can be finely tuned to provide ionic charge characteristics to facilitate uptake and distribution. Surprisingly, the present invention provides such compounds and compositions.
  • the present invention provides a variety of nanoparticles having a core portion and a surface of attached ligands, and optionally having attached co-ligand groups.
  • the core portion comprises gold or a gold alloy and the attached ligands can be a variety of saccharide or oligosaccharide components, at least a portion of which are C-glycosides.
  • a subject in need of treatment is administered a therapeutically effective amount of a composition comprising nanoparticles having a core portion and attached ligands, and optionally having attached co-ligands, wherein the core portion comprises gold or a gold alloy, and the attached ligands are the same or different, at least a portion of the attached ligands being C-glycosides that are capable of disrupting glycoprotein recognition.
  • Figure 1 illustrates the structures of alternate Gpl20 receptors including galactosyl ceramide (GalCer) and other glycosphingolipids.
  • Figure 2 provides a TEM image of Gal-Au (17, in (a)) and Glc-Au (18, in (b)). A drop of a solution of colloidal Au nanoparticles in nanopure water was deposited onto a silicon-coated copper grid and left to dry completely.
  • Figure 3 provides an AFM topograph of the Gal-Au (17) nanoparticles in (a) and a cursor profile along a selected line showing three particles with diameters of 2.0 nm, 1.6 nm and 2.1 nm, in (b).
  • Figure 4 provides the UN/Nis spectra of the Au nanoparticles.
  • FIG. 5 provides an illustration of the competitive B ⁇ AA competitive assay employed to characterize the activity of the Au nanoparticles.
  • Biotinylated GalCer is immobilized through interactions with ⁇ eutrAvidinTM and HRP-rgpl20 is introduced.
  • HRP-rgpl20 is competed off by the introduction of an alternate ligand. The extent to which the ligand is able to compete can be measured by comparing absorbance data from competition wells relative to bGalCer wells where no external ligand was introduced.
  • Figure 6 is a bar graph illustrating the results for the BNAA competition assay described in Example 4. Assay data is provided for disulfides 14, 15, and 16; Au nanoparticles 17, 18, and 19; and bGalCer 20 shown as endpoint readings at 415 nm after 20 min. As control, data for 20 correspond to maximal absorbance (no exposure to ligands in competition step).
  • Figure 7 illustrates the structures of the linking groups and mixed glyconanoparticles prepared according to Example 5.
  • treat refers to a method of alleviating or abrogating a disease and/or its attendant symptoms.
  • terapéuticaally effective amount refers to that amount of the compound being administered sufficient to prevent development of or alleviate to some extent one or more of the symptoms of the condition or disorder being treated.
  • composition as used herein is intended to encompass a product comprising the specified ingredients in the specified amounts, as well as any product which results, directly or indirectly, from combination of the specified ingredients in the specified amounts.
  • pharmaceutically acceptable it is meant the carrier, diluent or excipient must be compatible with the other ingredients of the formulation and not deleterious to the subject recipient thereof.
  • the "subject” is defined herein to include animals such as mammals, including, but not limited to, primates (e.g., humans), cows, sheep, goats, horses, dogs, cats, rabbits, rats, mice and the like. In preferred embodiments, the subject is a human.
  • nanoparticle compositions and methods are needed, and are provided herein, in which the polyvalent presentations are made with stable linkages. Additionally, the nanoparticles are preferably prepared by methods that allow the introduction of multiple ligands or co-ligands .
  • nanoparticles comprising a core portion and attached ligands, and optionally having attached co-ligands, wherein said core portion comprises gold or a gold alloy, and said attached ligands are the same or different, at least a portion of said attached ligands being C-glycosides.
  • the core will generally comprise gold, a gold alloy, or silica (i.e., SiO 2 ).
  • Suitable gold alloys include those made with silver, copper, iron, platinum, palladium and combinations thereof, particularly, silver and copper as well as silver, copper and palladium combinations.
  • the core portion is at least 80% gold, more preferably at least 95% gold, and still more preferably at least 99% gold.
  • the core of larger nanoparticles (e.g., greater than about 50 nm) comprises silica.
  • Suitable silica core portions for use in the present invention include those that are commercially available from Nissan Chemical industries under the name Snowtex and those that are made according to methods known in the art, such as those described in Stober et al, J. Colloid and Interface Science, 26:62-69 (1968).
  • the silica core portion is coated with gold or a gold alloy according to methods known in the art, such as those described in Pham et al, Langmuir, 18:4915-4920 (2002); Westcott et al, Langmuir, 14:5396-5401 (1998); and Oldenburg et al, Chemical Physics Letters, 288:243-247 (1998).
  • a 100 nm silica core portion can be coated with a 10 nm to 20 nm layer of gold or gold alloy.
  • the mean diameter of the core portion ranges from about 0.2 nm to about 150 nm so that the overall nanoparticle size has a mean particle diameter of from about 0.25 nm to about 160 nm, preferably from about 0.5 nm to 100 nm, more preferably from about 0.5 nm to about 5.0 nm, and still more preferably from about 1.0 nm to about 3.0 nm.
  • the mean diameter of the core portion is that which is comparable to the binding partner for the nanoparticle ligands.
  • the glycoprotein gpl20 has a size of about 2.5 nm and preferably binds to gold nanoparticles having a mean diameter of about 2.5 ⁇ 0.5 nm.
  • linking group or spacer generally having from 6 to 50 main chain atoms selected from the group consisting of S, O, N and C (hydrogen atoms will typically fill the remaining valences).
  • the spacer is selected to impart a variety of characteristics to the nanoparticle, such as increased stability, water solubility and circulation half-life.
  • the spacer is generally selected to reduce degradation or removal of the ligand from the core portion by being stable to hydrolytic processes (both enzymatic and chemolytic) under physiological conditions.
  • certain linking groups or spacers will have attached functional groups that operate to increase solubility (e.g., hydroxy, amino or ammonium groups).
  • the linking group or spacer portions are preferably of sufficient length to permit the ligand, or ligands, in the completed nanoparticle to interact freely with a target.
  • the spacer or linking group has a core- or surface-attaching portion and a longer chain portion.
  • the surface attaching portion is that part of the linking group which is directly attached to the core (e.g., Au).
  • Preferred attaching portions are thiols that are produced just prior to reaction with a gold or gold alloy core.
  • the longer chain portions are typically alkanes, alkenes, glycols (e.g., ethylene glycol oligomers containing multiple monomer units), alcohols, amines, polyamines (e.g., spermine, spermidine and polymeric derivatives thereof) or combinations thereof.
  • the linking group will be a polyethyleneglycol group which is at least a di or tri-ethyleneglycol.
  • the longer chain portion can be selected based upon its hydrophilic/hydrophobic properties to improve presentation of the ligands to certain receptors or proteins.
  • Attached to the distal portion of the linking group is a ligand or mixture of ligands that can be the same or different, with at least a portion of the attached ligands being C- glycosides.
  • the preparation of C-glycosides are known to those of skill in the art and exemplary processes are provided in detail below (see, Examples and Scheme 1).
  • C-glycosides can be used as ligands as monosaccharides, disaccharides, oligo- or polysaccharides.
  • C-glycosides are selected from C- or ⁇ , D or L, glycobiosyl alkylamines (e.g., meliobiosyl, lactobiosyl, and cellobiosyl); and longer oligosaccharides containing any combination of carbohydrate linkages involving C, N, O and S and combinations thereof.
  • Particularly preferred C-glycosides are selected from C-(j8-D- glycopyranosyl)methylamines and C-(3-D-galactopyranosyl)methylamines.
  • the term "glycoconjugate" refers to a conjugate between a C-glycoside ligand and a linking group.
  • the ligands are generally present on the surface of the core portion in an amount that optimizes binding with the target and reduces steric crowding of the ligands with one another.
  • the core portion has from 10 to 500 attached ligands, more preferably from 20 to 200 attached ligands, and still more preferably from 50 to 150 attached ligands.
  • the nanoparticles will further comprise attached co-ligands.
  • the te ⁇ n "co-ligands" refers to a moiety that facilitates ligand binding to a target by providing an environment that is conducive to such binding.
  • the co-ligands can be positively charged, negatively charged, or neutral.
  • a co-ligand will, in some embodiments, be a functional group that is positively charged at physiological pH and promotes solubility in physiological systems.
  • the co-ligand can provide a co-factor that facilitates target-ligand binding.
  • the co- ligand can be a targeting agent.
  • a variety of targeting agents are useful in the nanoparticles described herein.
  • the targeting agents are attached to the core portion in the same manner as described for the ligands, although in some instances, the linkages can be less robust than those used for the ligands.
  • the targeting agents themselves can be any element that makes it possible to direct the binding of a ligand on the nanoparticle to a particular site.
  • the targeting agent can be an extracellular targeting agent, which allows, for example, a nanoparticle to be directed towards certain types of cells or certain desired tissues (tumor cells, liver cells, hematopoietic cells, and the like).
  • the targeting agent can be a fusogenic peptide (e.g., gp41) for preventing cellular infection or a host cell fusogenic peptide for promoting cellular transfections, favoring the passage of the nanoparticle across membranes.
  • the targeting agent can also be a cell receptor ligand for a receptor that is present at the surface of the cell type, such as, for example, a carbohydrate, transferrin or insulin.
  • targeting agents useful in the context of the invention include peptides, hormones, vitamins, cytokines, oligonucleotides, lipids or sequences or fractions derived from these elements and which allow specific binding with their corresponding receptors.
  • the targeting agents are 5 sugars and/or peptides such as antibodies or antibody fragments, cell receptor ligands or fragments thereof, receptors or receptor fragments, and the like. More preferably, the targeting agents are ligands of growth factor receptors, of cytokine receptors, or of cell lectin receptors or of adhesion protein receptors.
  • the targeting agent can also be an antibody Fab fragment which makes it possible to target the Fc fragment receptor of immunoglobulins.
  • all of the linking groups attached to the core portion have a ligand attached thereto, e.g., only glycoconjugates are attached to the core portion.
  • a mixture of linking groups with attached ligands and linking groups without attached ligands e.g., a mixture of glycoconjugates and linking groups, are attached to the core portion. Suitable ratios of the linking groups with attached ligands to the linking groups
  • preparation of the nanoparticles of the present invention can be carried out using a variety of techniques known to the skilled synthetic chemist.
  • preparation of the nanoparticles begins with preparation of a suitable ligand having an attached linking group
  • C-glycosides which terminates in a functional group or protected functional group suitable (after removal of the protecting group) for attachment to the core portion.
  • the ligands used in the present invention are C-glycosides, selected for their stability in in vivo systems.
  • the preparation of C-glycosides can be accomplished by a variety of methods. Schemes 1-4, below, illustrate and describe one method for C-glycoside preparation, followed by
  • compositions comprising the nanoparticles described above in admixture with a pharmaceutically acceptable carrier or excipient.
  • the compositions are suitable for pharmaceutical or diagnostic use.
  • the invention provides the subject nanoparticles combined with a pharmaceutically acceptable excipient such as sterile saline or other medium, water, gelatin, an oil, etc. to form pharmaceutically acceptable compositions.
  • a pharmaceutically acceptable excipient such as sterile saline or other medium, water, gelatin, an oil, etc.
  • the compositions can be administered alone or in combination with another convenient carrier, diluent, etc. and such administration may be provided in single or multiple dosages.
  • Useful carriers include solid, semi-solid or liquid media including water and non-toxic organic solvents.
  • compositions can be provided in any convenient form, including tablets, capsules, lozenges, troches, hard candies, powders, sprays, creams, suppositories, etc.
  • the compositions in pharmaceutically acceptable dosage units or in bulk, may be incorporated into a wide variety of containers.
  • dosage units may be included in a variety of containers including capsules, pills, etc.
  • the present invention provides methods for the use of the foregoing nanoparticles and compositions.
  • the invention provides methods for treating diseases dependent on glycoprotein recognition. The methods typically involve administering to a patient an effective formulation of one or more of the subject nanoparticles.
  • the invention provides methods of using the subject nanoparticles and compositions to treat disease or provide medicinal prophylaxis to individuals who possess a compromised immune system or are expected to suffer immunosuppressed conditions, such as patients prior to undergoing immunosuppressive therapy in connection with organ transplantation or anticancer chemotherapy. These methods generally involve administering to the host an effective amount of the subject compounds or pharmaceutically acceptable compositions.
  • the nanoparticles of the present invention are advantageously combined and/or used in combination with other antiviral agents useful in the treatment and/or prevention of the viral infections described herein.
  • the nanoparticles may be advantageously combined and/or used in combination with agents useful in the treatment and/or prevention of conditions often associated with the viral infections described herein, such as anti-HIN agents (described below), immunostimulatory agents (e.g., vaccines) or immunosuppressive agents (e.g., cyclosporin, FK-506 and rapamycin).
  • anti-HIN agents described below
  • immunostimulatory agents e.g., vaccines
  • immunosuppressive agents e.g., cyclosporin, FK-506 and rapamycin.
  • administration of the subject compounds or compositions in conjunction with these alternative agents enhances the efficacy of such agents.
  • the present nanoparticles, when combined or administered in combination with other antiviral or immunosuppressive agents can be used in dosages which are less than the expected amounts
  • Suitable agents for combination therapy include those that are currently commercially available and those that are in development or will be developed. While antiviral agents may be particularly suitable for the treatment or prevention of a particular viral disorders), practitioners skilled in the art understand that such agents frequently are useful in treating a range of viral-related disorders. Exemplary agents useful in the treatment of certain viral infections include acyclovir, cidofovir, ganciclovir, immunoglobuhn and foscarnet.
  • nucleoside/nucleotide analogs valaciclovir, valganciclovir, adefovir, dipivoxil and lobucavir include the nucleoside/nucleotide analogs valaciclovir, valganciclovir, adefovir, dipivoxil and lobucavir; the antisense agents fomivirsen, GEM 132 (Hybridon), ISIS 13312 (ISIS); and other therapies like benzimidavir and sevirumab.
  • Exemplary anti-HJN agents include nucleoside analog reverse transcriptase inhibitors such as zidovudine (AZT), didanosine (ddl), zalcitabine (ddC, dideoxycytidine), stavudine (d4T), lamivudine (3TC), abacavir (1592U89), emtricitabine (FTC, Triangle Pharmaceuticals), BCH-10652 (BioChem Pharma) and the related nucleotide analogs (e.g., PMPA (Gilead Sciences)); non-nucleoside reverse transcriptase inhibitors such as nevirapine (NVP), delavirdine (DLV), efavirenz (DMP-266), emivirine (MKC-442), AG1549 (Agouron Pharmaceuticals; PNU142721 (Pharmacia),calanolide-A (Sarawak MediChem Pharmaceuticals); protease inhibitors such as saquinavir (
  • anti-HIN agents that may be used in combination with the compounds and compositions of the present invention include HIN integrase inhibitors (e.g., AR-177 (Aronex Pharmaceuticals)), fusion inhibitors (e.g., T-20 (Roche)) and antisense drugs (e.g., HGTN43 (Enzo Therapeutics)).
  • HIN integrase inhibitors e.g., AR-177 (Aronex Pharmaceuticals)
  • fusion inhibitors e.g., T-20 (Roche)
  • antisense drugs e.g., HGTN43 (Enzo Therapeutics)
  • nanoparticles and compositions of the present invention may also be advantageously used as antiviral prophylactic treatment in combination with immunosuppressive protocols such as bone-marrow destruction (either by radiation or chemotherapy).
  • compositions and nanoparticles of the invention and the pharmaceutically acceptable salts thereof can be administered in any effective way such as via oral, parenteral or topical routes.
  • the compounds are administered in dosages ranging from about 2 mg up to about 2,000 mg per day, although variations will necessarily occur depending on the disease target, the patient, and the route of administration.
  • Preferred dosages are administered orally in the range of about 0.05 g/kg to about 20 mg/kg, more preferably in the range of about 0.05 mg/kg to about 2 mg/kg, most preferably in the range of about 0.05 mg/kg to about 0.2 mg per kg of body weight per day.
  • a home-built scanner with the optical beam deflection configuration was used for atomic force microscopy (AFM) studies. See, Liu, G. Y.; Fenter, P.; Chidsey, C. E. D.; Ogletree, D. F.; Evenberger, P.; Salmeron, M. Chem. Phys. 1994, 101, 4301.
  • the electronic controllers and software are commercially available from RHK Technology, Inc. (Troy, MI).
  • the AFM scanner was calibrated using both mica(OOOl) and Au(l 11) surfaces. Sharpened Si 3 N 4 microlevers (ThermoMicroscopes) with a force constant of 0.1 N/m were used for AFM imaging. Images were acquired in sec-butanol, and the typical imaging force was 0.1 nN. Nanoparticles were deposited onto atomically flat mica(0001) surfaces which were prepared by freshly cleaving them prior to nanoparticle deposition.
  • the resin was filtered off and washed with MeOH and CH 2 C1 2 , the filtrate was concentrated and the resulting material was purified by column chromatography (eluent: CH 2 Cl 2 /MeOH v/v 10:1 to 5:1).
  • the desired compound (10) (153 mg, 0.38 mmol) was obtained in 74% yield as a colorless foam.
  • Glyconanoparticles Gal-Au (17, 13 mg, 88%), Glc-Au (18, 14 mg, 93%), and Sp-Au (19, 13 mg, 91%) were obtained as dark-brown solids. The yields were calculated based upon Au as the limiting reagent.
  • Gal-Au (17) 1H NMR (500 MHz, D 2 O) ⁇ 3.30 - 3.43 (bm, 3 H, H-2', H-l', H-3'), 3.55 - 3.79 (bm, 13 H, H-4', H-6', H-7', H-7", H-l ", H-3, H-4, H-5, H-6 (weak)), 3.87 (bs, 1 H, H-5'), 4.06 (bs, 2 H, H-2).
  • This example illustrates methods for the characterization of the Au nanoparticles.
  • NMR and IR Characterization of the Au Nanoparticles Evidence for incorporation of glycoconjugates onto the Au nanoparticles was provided by 1H NMR spectra of Gal-Au (17) and Glc-Au (18) (ca. 5 mg per 0.8 mL D 2 O), which showed typical broadening characteristic of mono-layer protected gold clusters. See, for example, Terrill, R. H.; Postlethwhaite, T. A.; Chen, C.-H.; Poon, C.-D.; Terzis, A.; Chen, A.; Hutchison, J. E.; Clark, M. R; Wignall, G. D.; Londono, J.
  • Figure 3 a represents a typical AFM topograph of the Gal-Au (17) nanoparticles.
  • the height measured from the cursor profiles in the AFM topographic images was utilized to quantify the particle diameter.
  • the cursor profile in Figure 3b shows the diameters of the three particles are 2.0 nm, 1.6 nm and 2.1 nm, respectively. With 162 nanoparticles imaged, the measured height, i.e. the particle diameter ranged from 1.2 to 4.2 nm.
  • SP band surface plasmon band
  • the SP band intensity correlates with core size (Nezmar, I.; Alvarez, M. M.; Khoury, J. T.; Salisbury, B. E.; Shafigullin, M. ⁇ .; Whetten, R. L. Z. Phys D 1997, 401, 147-151; Schaaf, T. G.; Shafigullin, M. N.; Khoury, J. T.; Nezmar, I; Whetten, R. L.; Cullen, W. G.; First, P. ⁇ .; Wing, C; Ascensio, J.; Yacaman, M. J.
  • Biotin ⁇ eutrAvidinTM Adhesion Assays (B ⁇ AA): Recombinant gpl20 ⁇ ⁇ B (HJN- 1)-HRP was obtained from Bartels - A Trinity Biotech Company, Reacti-BindTM
  • Dried analyte 16 and freeze-dried analytes 14, 15, 17, 18, 19, and 20 were weighed out to the tenth of a microgram using a Satorius BL 120S balance.
  • Stock solutions were prepared at 1 mg/mL in 20% DMSO/80% PBS buffer. The solutions were thoroughly mixed prior to further dilution in PBS, and they were stored at -80 °C between uses.
  • the concentration of bGalCer as well as biotinamide (blocking agent) utilized in the assays was 20 ⁇ g/mL.
  • the plates were prepared by first incubating with bGalCer and biotinamide.
  • a solution of rgpl20-HRP was prepared before use by 1 :3 dilution of a 500 ⁇ L stock vial containing 0.333 ⁇ g rgpl20-HRP with PBS buffer and introduced into the wells.
  • analytes 14, 15, 16 were incubated at concentrations of 0.1, 1, 10, 100, 500, 1000 ⁇ g/mL and the Au nanoparticles 17, 18, 19 were assayed at concentrations of 1, 10, 20, 40, 80, 100, 200, 400 ⁇ g/mL on a JitterbugTM microplate incubator (Boekel).
  • the addition of rgpl20 was omitted and the contribution of Sp-Au (19) to background absorbance was monitored at each concentration.
  • the plate absorbances were read every min for 20 min at 630 nm (with 5 s shaking before each reading) using a Bio-Rad model 550 microplate reader. Each well reading had the TMB blank automatically subtracted through the use of the Biorad Microplate manager software. Endpoints were read at 415 nm after addition of 100 ⁇ L of 2 N H 2 SO 4 to each well. The results given for these assays are representative of several separate experiments.
  • the EC 50 value represents the concentration of the compound required to induce 50% displacement of rgpl20-HRP bound to bGalCer.
  • the EC 50 values were calculated from dose-response curves (rgpl20 bound to bGalCer in % vs. analyte cone.) plotted using exponential trendlines (Microsoft® Excel 2000). The error was determined using a transformed regression model (Microsoft® Excel 2000).
  • the calculated EC 50 values for Au glyconanoparticles include an additional error reflected in the elemental analysis (Gal-Au (17):1.5%; Glc-Au (18): 1.4%; and Sp-Au (19): 1.2%).
  • the Gal-S2 (14) required approximately 1 mg/mL to displace 50% of bound rgpl20 indicating that it was about twice as active as the Glc-S2 (15) analog.
  • Au nanoparticles functionalized with only the spacer acid (19) showed similar activity to the glycosylated disulfides (2.4 mg/mL). This activity is attributed to non-specific electrostatic interactions of the highly negatively charged surface (free acid functionality) with basic amino acids present in rgpl20. See, Rico-Lattes, I.; Gouzy, M.-F., Andre-Barres, C; Guidetti, B.; Lattes, A. New J. Chem. 1998, 22, 451.
  • Table 1 ECso values calculated from dose-response curves (graph: % rgpl20 bound to bGalCer vs. analyte cone.) plotted using exponential trendlines
  • the Au glyconanoparticles 17 and 18 exhibit significantly higher activity (>300X) when compared to the corresponding disulfides 14 and 15, respectively, even without factoring in the gold content.
  • Table 1 the molecular weight of Gal-Au and Glc-Au is 96,190 and 101,676 g/mol respectively, which correlates with mM EC50 values of 5.6 and 13.6 x 10-5, respectively.
  • Whitesides and co- workers have described a term ⁇ that relates mono- and polyvalency by accounting for the difference in the number of ligands presented to the protein in each case. See, Mammen, M.; Choi, S.K.; Whitesides, G.M., Angew.
  • the ⁇ values indicate that the galactosyl containing disulfide (14) has approximately twice the activity of the glucosyl disulfide (15), yet 14 is only 12% as active as biotinylated GalCer (20). This is not surprising, since several studies have shown that a lipid component is required for maximal affinity.
  • the relative activity between galactosyl and glucosyl head groups is maintained in the Au glyconanoparticles with 17 being approximately twice as active as 18. However, 17 is 450x more active than 14 (54 divided by 0.12) and 18 is > 300x more active than 15 (22 divided by 0.07).
  • the enhanced binding activity of the Au glyconanoparticles relative to the disulfides may be due to fundamental changes in the recognition process of divalent vs. polyvalent species resulting in formation of thermodynamically more stable polyvalent ligand/rgpl20 complexes.
  • kinetic factors reflecting differences in on/off rates of divalent and polyvalent ligands may account for the enhanced activity. See, Liang, R., Loebach, J., Horan, N., Ge, M., Thompson, C, Yan, L., Kahne, D., Pro Natil. Acad. Set., 1997, 94, 10554- 10559 and references therein.
  • This example illustrates the preparation of mixed glyconanoparticles, t.e., gold glyconanoparticles containing a mixture of a glycoconjugate and a linking group, wherein the glycoconjugate to linking group ratio is about 75:25, 50:50, or 25:75.
  • Number for the compounds prepared corresponds to the numbering in Figure 7.
  • WO 02/32404 A2 a freshly prepared 25 mM solution of hydrogen tetrachloroaurate (III) hydrate (0.167 mmol, 1 eq) in MeOH was added to a mixture of a freshly prepared 12 mM solution of one of the glycoconjugate disulfide derivatives (14, 15) and the disulfide derivative (23) (i.e., Sp-OH) at molar ratios of 75:25, 50:50, and 25:75 (total 0.25 mmol, total 1.5 eq) in MeOH. Under vigorous stirring, a freshly prepared 1 M solution of NaBH 4 (3.34 mmol, 20 eq) in MeOH was added drop wise. Under liberation of H 2 , dark- brown suspensions formed instantly.
  • the mixed glyconanoparticles Gal/Sp-COOH-Au (27) and Glc/Sp- COOH-Au (28) were prepared from a mixture of one of the glycoconjugate disulfide derivatives (14, 15) and the disulfide derivative (16) (i.e., Sp-COOH) at molar ratios of 75:25, 50:50, and 25:75 (total 0.25 mmol, total 1.5 eq) in MeOH.

Landscapes

  • Health & Medical Sciences (AREA)
  • Chemical & Material Sciences (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Medicinal Chemistry (AREA)
  • Pharmacology & Pharmacy (AREA)
  • General Health & Medical Sciences (AREA)
  • Epidemiology (AREA)
  • Animal Behavior & Ethology (AREA)
  • Public Health (AREA)
  • Veterinary Medicine (AREA)
  • Engineering & Computer Science (AREA)
  • Inorganic Chemistry (AREA)
  • Bioinformatics & Cheminformatics (AREA)
  • Molecular Biology (AREA)
  • Nanotechnology (AREA)
  • Biochemistry (AREA)
  • Ceramic Engineering (AREA)
  • Biophysics (AREA)
  • Biotechnology (AREA)
  • General Engineering & Computer Science (AREA)
  • Medical Informatics (AREA)
  • Crystallography & Structural Chemistry (AREA)
  • Immunology (AREA)
  • Pharmaceuticals Containing Other Organic And Inorganic Compounds (AREA)
  • Preventing Corrosion Or Incrustation Of Metals (AREA)

Abstract

L'invention concerne des nanoparticules comprenant une partie centrale en or ou en alliage d'or et une surface de ligands fixés et, éventuellement, des groupes de co-ligands fixés. Les ligands fixés peuvent être une variété de composants du saccharide ou de l'oligosaccharide, au moins une partie de ceux-ci étant des C-glycosides.
PCT/US2004/017607 2003-06-03 2004-06-02 Preparation et utilisation de glyconanoparticules en or Ceased WO2005010481A2 (fr)

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
US47583603P 2003-06-03 2003-06-03
US60/475,836 2003-06-03

Publications (2)

Publication Number Publication Date
WO2005010481A2 true WO2005010481A2 (fr) 2005-02-03
WO2005010481A3 WO2005010481A3 (fr) 2005-08-04

Family

ID=34102628

Family Applications (1)

Application Number Title Priority Date Filing Date
PCT/US2004/017607 Ceased WO2005010481A2 (fr) 2003-06-03 2004-06-02 Preparation et utilisation de glyconanoparticules en or

Country Status (1)

Country Link
WO (1) WO2005010481A2 (fr)

Family Cites Families (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US6530944B2 (en) * 2000-02-08 2003-03-11 Rice University Optically-active nanoparticles for use in therapeutic and diagnostic methods
GB0025414D0 (en) * 2000-10-16 2000-11-29 Consejo Superior Investigacion Nanoparticles

Also Published As

Publication number Publication date
WO2005010481A3 (fr) 2005-08-04

Similar Documents

Publication Publication Date Title
US20230132265A1 (en) Compounds for the prevention and treatment of diseases and the use thereof
US7579324B2 (en) Conjugates of biologically active compounds, methods for their preparation and use, formulation and pharmaceutical applications thereof
ES2346319T5 (es) Nanopartículas
US8357506B2 (en) Method of identifying improved conjugates of biologically active compounds
JP4367798B2 (ja) インビボ許容性の向上した製剤
CN111032670A (zh) 用于治疗系统性胰岛素抗性病症的化合物及其用途
KR102053065B1 (ko) 히알루론산 및 독소루비신을 이용한 pH 감응성 항암 엑소좀 조성물
EP1687024A2 (fr) Conjugues de nanoparticules-antigenes carbohydrate et utilisation comme agents anti-metastatiques dans le traitement du cancer
JP2005508332A (ja) 薬剤を送達するための、輸送タンパク質とコンジュゲートコバラミンとの同時投与
US7531162B2 (en) Transcobalamin receptor binding conjugates useful for treating abnormal cellular proliferation
WO2005051429A2 (fr) Systeme d'administration ciblee pour agents bioactifs
KR20010006280A (ko) 다수개의 활성 잔기를 제공하는 분자
US20050026996A1 (en) Taxane prodrugs
CN109071585B (zh) 预防和治疗半乳糖凝集素相关疾病的硒代半乳糖苷化合物及其用途
Farabi et al. Concise and Reliable Syntheses of Glycodendrimers via Self-Activating Click Chemistry: A Robust Strategy for Mimicking Multivalent Glycan–Pathogen Interactions
US20130012466A1 (en) Colchicine neoglycosides and methods for their synthesis and use
WO2002094869A1 (fr) Proteine lectine preparee a partir de maackia fauriei, procede de preparation et utilisation de cette proteine
JP2026502084A (ja) 脂質化合物及び脂質ナノ粒子組成物
JP2022176381A (ja) 乳がんまたは胃がんの治療のための抗体－薬物複合体
WO2005010481A2 (fr) Preparation et utilisation de glyconanoparticules en or
WO2009095480A1 (fr) Traitements antitumoraux améliorés
JPWO2007018272A1 (ja) 糖鎖修飾リポソーム及び該リポソームを含有する薬剤送達組成物
CN110393797A (zh) 基于糖脂佐剂的糖肽疫苗的制备方法与应用
JP2022546415A (ja) 治療薬の核酸媒介性送達
Dai et al. A self-assembling conjugate of SN38 with aminoguanidine for simultaneously suppressing proliferation and migration of breast cancer cells

Legal Events

Date Code Title Description
AK Designated states

Kind code of ref document: A2

Designated state(s): AE AG AL AM AT AU AZ BA BB BG BR BW BY BZ CA CH CN CO CR CU CZ DE DK DM DZ EC EE EG ES FI GB GD GE GH GM HR HU ID IL IN IS JP KE KG KP KR KZ LC LK LR LS LT LU LV MA MD MG MK MN MW MX MZ NA NI NO NZ OM PG PH PL PT RO RU SC SD SE SG SK SL SY TJ TM TN TR TT TZ UA UG US UZ VC VN YU ZA ZM ZW

AL Designated countries for regional patents

Kind code of ref document: A2

Designated state(s): GM KE LS MW MZ NA SD SL SZ TZ UG ZM ZW AM AZ BY KG KZ MD RU TJ TM AT BE BG CH CY CZ DE DK EE ES FI FR GB GR HU IE IT LU MC NL PL PT RO SE SI SK TR BF BJ CF CG CI CM GA GN GQ GW ML MR NE SN TD TG

121 Ep: the epo has been informed by wipo that ep was designated in this application
122 Ep: pct application non-entry in european phase