[go: up one dir, main page]

EP1885335A1 - Microparticules a surface modifiee et procedes de formation et d'utilisation associes - Google Patents

Microparticules a surface modifiee et procedes de formation et d'utilisation associes

Info

Publication number
EP1885335A1
EP1885335A1 EP06751564A EP06751564A EP1885335A1 EP 1885335 A1 EP1885335 A1 EP 1885335A1 EP 06751564 A EP06751564 A EP 06751564A EP 06751564 A EP06751564 A EP 06751564A EP 1885335 A1 EP1885335 A1 EP 1885335A1
Authority
EP
European Patent Office
Prior art keywords
microparticle
monolayer
charged
preformed
active agent
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.)
Withdrawn
Application number
EP06751564A
Other languages
German (de)
English (en)
Inventor
Julia Rashba-Step
Terrence L. Scott
Ramin Darvi
Yuri Lvov
Tatsiana Shutava
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.)
Baxter Healthcare SA
Baxter International Inc
Louisiana Tech University
Original Assignee
Baxter Healthcare SA
Baxter International Inc
Louisiana Tech University
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 Baxter Healthcare SA, Baxter International Inc, Louisiana Tech University filed Critical Baxter Healthcare SA
Publication of EP1885335A1 publication Critical patent/EP1885335A1/fr
Withdrawn legal-status Critical Current

Links

Classifications

    • 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
    • A61K31/7088—Compounds having three or more nucleosides or nucleotides
    • A61K31/7105—Natural ribonucleic acids, i.e. containing only riboses attached to adenine, guanine, cytosine or uracil and having 3'-5' phosphodiester links
    • 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
    • A61K9/0012—Galenical forms characterised by the site of application
    • A61K9/0019—Injectable compositions; Intramuscular, intravenous, arterial, subcutaneous administration; Compositions to be administered through the skin in an invasive manner
    • 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
    • A61K9/0012—Galenical forms characterised by the site of application
    • A61K9/0019—Injectable compositions; Intramuscular, intravenous, arterial, subcutaneous administration; Compositions to be administered through the skin in an invasive manner
    • A61K9/0024—Solid, semi-solid or solidifying implants, which are implanted or injected in body tissue
    • 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
    • A61K9/10—Dispersions; Emulsions
    • A61K9/127—Synthetic bilayered vehicles, e.g. liposomes or liposomes with cholesterol as the only non-phosphatidyl surfactant
    • A61K9/1271—Non-conventional liposomes, e.g. PEGylated liposomes or liposomes coated or grafted with polymers
    • A61K9/1272—Non-conventional liposomes, e.g. PEGylated liposomes or liposomes coated or grafted with polymers comprising non-phosphatidyl surfactants as bilayer-forming substances, e.g. cationic lipids or non-phosphatidyl liposomes coated or grafted with polymers
    • 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
    • A61K9/14—Particulate form, e.g. powders, Processes for size reducing of pure drugs or the resulting products, Pure drug nanoparticles
    • A61K9/16—Agglomerates; Granulates; Microbeadlets ; Microspheres; Pellets; Solid products obtained by spray drying, spray freeze drying, spray congealing,(multiple) emulsion solvent evaporation or extraction
    • 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
    • A61K9/14—Particulate form, e.g. powders, Processes for size reducing of pure drugs or the resulting products, Pure drug nanoparticles
    • A61K9/16—Agglomerates; Granulates; Microbeadlets ; Microspheres; Pellets; Solid products obtained by spray drying, spray freeze drying, spray congealing,(multiple) emulsion solvent evaporation or extraction
    • A61K9/1605—Excipients; Inactive ingredients
    • A61K9/1629—Organic macromolecular compounds
    • A61K9/1635—Organic macromolecular compounds obtained by reactions only involving carbon-to-carbon unsaturated bonds, e.g. polyvinyl pyrrolidone, poly(meth)acrylates
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K9/00—Medicinal preparations characterised by special physical form
    • A61K9/14—Particulate form, e.g. powders, Processes for size reducing of pure drugs or the resulting products, Pure drug nanoparticles
    • A61K9/16—Agglomerates; Granulates; Microbeadlets ; Microspheres; Pellets; Solid products obtained by spray drying, spray freeze drying, spray congealing,(multiple) emulsion solvent evaporation or extraction
    • A61K9/1605—Excipients; Inactive ingredients
    • A61K9/1629—Organic macromolecular compounds
    • A61K9/1658—Proteins, e.g. albumin, gelatin
    • 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
    • A61K9/14—Particulate form, e.g. powders, Processes for size reducing of pure drugs or the resulting products, Pure drug nanoparticles
    • A61K9/16—Agglomerates; Granulates; Microbeadlets ; Microspheres; Pellets; Solid products obtained by spray drying, spray freeze drying, spray congealing,(multiple) emulsion solvent evaporation or extraction
    • A61K9/1682—Processes
    • A61K9/1688—Processes resulting in pure drug agglomerate optionally containing up to 5% of excipient
    • 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
    • A61K9/5005—Wall or coating material
    • A61K9/5021—Organic macromolecular compounds
    • A61K9/5026—Organic macromolecular compounds obtained by reactions only involving carbon-to-carbon unsaturated bonds, e.g. polyvinyl pyrrolidone, poly(meth)acrylates
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K9/00—Medicinal preparations characterised by special physical form
    • A61K9/48—Preparations in capsules, e.g. of gelatin, of chocolate
    • A61K9/50—Microcapsules having a gas, liquid or semi-solid filling; Solid microparticles or pellets surrounded by a distinct coating layer, e.g. coated microspheres, coated drug crystals
    • A61K9/5005—Wall or coating material
    • A61K9/5021—Organic macromolecular compounds
    • A61K9/5036—Polysaccharides, e.g. gums, alginate; Cyclodextrin
    • 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
    • A61K9/5005—Wall or coating material
    • A61K9/5021—Organic macromolecular compounds
    • A61K9/5036—Polysaccharides, e.g. gums, alginate; Cyclodextrin
    • A61K9/5042—Cellulose; Cellulose derivatives, e.g. phthalate or acetate succinate esters of hydroxypropyl methylcellulose
    • A61K9/5047—Cellulose ethers containing no ester groups, e.g. hydroxypropyl methylcellulose
    • 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
    • A61K9/5005—Wall or coating material
    • A61K9/5021—Organic macromolecular compounds
    • A61K9/5052—Proteins, e.g. albumin
    • 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
    • A61K9/5005—Wall or coating material
    • A61K9/5021—Organic macromolecular compounds
    • A61K9/5052—Proteins, e.g. albumin
    • A61K9/5057—Gelatin
    • 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
    • A61K9/5073—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 having two or more different coatings optionally including drug-containing subcoatings
    • 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
    • A61K9/5089—Processes
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P5/00—Drugs for disorders of the endocrine system
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P5/00—Drugs for disorders of the endocrine system
    • A61P5/48—Drugs for disorders of the endocrine system of the pancreatic hormones
    • A61P5/50—Drugs for disorders of the endocrine system of the pancreatic hormones for increasing or potentiating the activity of insulin
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K17/00—Carrier-bound or immobilised peptides; Preparation thereof
    • C07K17/02—Peptides being immobilised on, or in, an organic carrier
    • C07K17/08—Peptides being immobilised on, or in, an organic carrier the carrier being a synthetic polymer

Definitions

  • the present disclosure is generally directed to microparticles containing one or more active agents and delivery methods of such microparticles to subjects. More particularly, the present disclosure is directed to surface modifications of the microparticles such that they are capable of controlled release of one or more active agents. The present disclosure is further directed to methods for making and using such surface-modified microparticles.
  • Microparticles have been used in many different applications, including the controlled delivery and/or release of active agents. Controlling or modifying the release profile of an active agent can, if desired, prolong the levels (such as therapeutic levels) of the active agent in the blood stream of the recipient, improve pharmacokinetics and pharmacodynamics, and result in greater convenience to the recipient.
  • the present disclosure is generally directed to methods for preparing a surface-modified microparticle.
  • the method includes providing an amorphous and solid preformed microparticle including at least one active agent.
  • the preformed microparticle has an outer surface carrying a net surface charge.
  • the method further includes exposing at least the outer surface of the preformed microparticle to at least one charged compound having a net charge that is opposite in sign to the net surface charge of the preformed microparticle outer surface.
  • a monolayer of the charged compound is formed whereby the monolayer is associated with the preformed microparticle outer surface.
  • the present disclosure is also directed to a method for preparing a surface- modified microparticle including providing a liquid continuous phase system containing a solvent, at least one active agent, and one or more phase-separation enhancing agents.
  • the method includes inducing a liquid-solid phase separation at, optionally, a controlled rate to cause a liquid-solid separation, and forming a solid phase that includes a solid and amorphous microparticle containing the active agent, the microparticle having an outer surface carrying a net surface charge, while the solvent and the phase-separation enhancing agents remain in the liquid phase.
  • the method further includes forming a monolayer including at least one charged compound on the formed microparticle whereby the formed monolayer is associated with the microparticle outer surface.
  • the present disclosure is also directed to a method for preparing a surface modified microparticle that includes providing an amorphous and solid preformed microparticle including at least one active agent, the preformed microparticle having an outer surface carrying a net surface charge.
  • the method further includes exposing at least the outer surface of the preformed microparticle to at least one charged compound having a net charge that is opposite in sign to the net surface charge of the preformed microparticle.
  • An intermediate microparticle is formed that includes the preformed microparticle and a formed monolayer including the at least one charged compound wherein the formed monolayer is associated with the preformed microparticle outer surface.
  • the formed monolayer is then exposed to at least a different charged compound to form a surface modified microparticle that includes the intermediate microparticle and a subsequent monolayer including at least the one different charged compound.
  • the surface modified microparticle has a release profile for release of the at least one active agent that is different from the release profile of the intermediate microparticle.
  • the present disclosure is also directed to a microparticle that includes a solid, amorphous core microparticle that includes between 80% or greater than by weight, of at least one active agent.
  • the outer surface of the core microparticle carries a selected net surface charge.
  • a monolayer of at least one charged compound carrying a net surface charge that is sufficiently different from the surface charge of the core microparticle outer surface to allow for association therewith, is associated with the core microparticle outer surface at least by, but not limited to, electrostatic interaction with the outer surface.
  • a solid microparticle including at least 80%, by weight, of at least one active agent where the outer surface of the microparticle includes at least one charged compound associated with the active agent may display a 1-hour percentage of cumulative release of the active agent that is 50% or less when subjected to in vitro release in a suitable buffer at selected pH and temperature.
  • a solid, preformed microparticle having at least 80%, by weight, of at least one proteinaceous compound where the outer surface of the preformed microparticle has at least one charged compound associated with the proteinaceous compound are suitable for in vivo administration.
  • the microparticle Upon such administration, the microparticle provides a C max and t max that is different from the C max and t max of the core microparticle.
  • microparticles [00010] Further details of the above-described microparticles, methods of making microparticles and methods for controlling the release of active agents from the microparticles are discussed below.
  • FIG. 1 is a flow chart showing an exemplary method set forth in the present disclosure
  • FIG. 2 is a schematic illustration of the fabrication of monolayers of charged compounds on a preformed microparticle
  • Fig. 3 is a graph that shows the change in zeta potential of insulin microparticles with alternatingly charged monolayers (Examples IA & IB);
  • Fig. 4 is a graph that shows the change in zeta potential of insulin microparticles, each, with one monolayer of different polyanionic compounds (Example 2A);
  • Fig. 5 is a graph that shows the change in zeta potential of insulin microparticles, each with one monolayer of different polycationic compounds (Example 2B);
  • Fig. 6 is a laser scanning confocal (LSC) micrograph of insulin microparticles with one FITC-labeled protamine monolayer (Example 2B);
  • Fig. 7 is a graph showing surface electric charge alteration of insulin microparticles, each with a first monolayer of different polyanionic compounds and a second monolayer of poly-L-lysine (Example 3A);
  • Fig. 8 is a LSC micrograph of insulin microparticles with a first monolayer of PSS and a second monolayer of FITC- labeled PLL (Example 3A);
  • Fig. 9 is a graph showing progressive film thicknesses after each successive deposition of alternatingly charged polyelectrolyte monolayers (e.g., PLL and chondroitin sulfate) on the QCM electrode (Example 3A);
  • alternatingly charged polyelectrolyte monolayers e.g., PLL and chondroitin sulfate
  • Fig. 10 is a graph showing changes in zeta-potential of insulin microparticles following subsequent depositions of a chondroitin sulfate monolayer and a gelatin A monolayer (Example 3B);
  • Fig. 11 is a graph comparing zeta-potential of insulin microparticles with monolayers of protamine sulfate and chondroitin sulfate in the presence and absence of zinc cations in the PEG-containing reaction medium (Example 4A);
  • Fig. 12 is a graph showing zeta-potential of insulin microparticles with monolayers of protamine sulfate and chondroitin sulfate in presence of zinc cations in a
  • Fig. 13 is a graph showing LSC micrograph of insulin microparticles with one monolayer of Rodamine B-labeled protamine (top left) and one monolayer of FITC- labeled DAP (top right) (Example 5);
  • Fig. 14 is a graph showing the zeta-potential of insulin microparticles following deposition of each monolayer as described in Example 5;
  • Fig. 15 is a graph showing insulin release profiles from microparticles coated with a monolayer of protamine sulfate with various concentrations of the polycation in the reaction medium (Example 6);
  • Fig. 16 is a graph showing insulin release profiles from microparticles with a first monolayer of protamine sulfate and a second monolayer of carboxymethyl cellulose at various concentrations of the polyanion in the reaction medium (Example 7);
  • Fig. 17 is a graph showing release profiles of insulin from insulin microparticles coated with three monolayers (Example 7);
  • Fig. 18A is a graph showing the serum human insulin (hINS) concentration versus time profiles in rats that have received a single subcutaneous injection of uncoated insulin microparticles, or protamine-coated insulin microparticles (Example 8);
  • Fig. 18B is a graph showing the serum glucose depression versus time profiles of rats treated with single subcutaneous injection of uncoated insulin microparticles, or protamine-coated insulin microparticles Example 8);
  • Fig. 19 is a graph showing alteration in the surface charge of insulin microparticle by deposition of one monolayer of protamine in various solubility-reducing media at pH 7.0 (Example 9);
  • Fig. 2OA is a graph showing the effect of protamine concentration in the reaction medium on the surface charge of insulin microparticles, and the extent of the dissolution after 48 h from initiation of the in vitro release (Example 10);
  • Fig. 2OB is a graph showing the effect of CMC concentration in the reaction medium on the surface charge of protaimne-coated insulin microparticles, and the extent of their dissolution after 48 h from initiation of the in vitro release (Example 10);
  • Fig. 2OC is a graph showing the effect of protamine concentration in the reaction medium on the surface charge of insulin microparticles coated with one monolayer of protamine and one monolayer of CMC, and the extent of their dissolution after 48 h from initiation of the in vitro release (Example 10);
  • Fig. 21 A is a graph showing the alteration of zeta-potential of hGH microparticles following subsequent depositions of protamine sulfate and chondroitin sulfate monolayers (Example 11);
  • Fig. 21B is a graph showing profiles of hGH release from microparticles coated with one, two, or three monolayers of alternating protamine and condroitin sulfate
  • Fig. 22 is a graph showing the alteration of zeta-potential of intravenous immunoglobulin (IVIG) microparticles following subsequent depositions of chondroitin sulfate and protamine sulfate monolayers (Example 12);
  • Fig. 23 is a graph showing net surface charge characteristics of insulin microspheres in 16% PEG solution across a pH range of 4 to 7.5 (Example 13);
  • Fig. 24 is a flow chart showing another exemplary method of preparing microparticles set forth in the present disclosure.
  • Fig. 25 is a graph showing the effect of reaction pH on the zeta potential of insulin microparticles that have been surface-modified with one monolayer of protamine sulfate, poly-1-lysine, or poly-1-arginine (Example 14);
  • Fig. 26 is a graph showing the effect of reaction pH on the in vitro 1-hour percentage of cumulative insulin release of insulin microparticles surface-modified with one monolayer of protamine sulfate, poly-1-lysine, or poly-1-arginine (Example 14);
  • Fig. 27 is a laser scanning confocal (LSC) micrograph of nucleic acid microparticles surface-modified with one monolayer of rodamine B-labeled poly-1-lysine (Example 15);
  • Fig. 28 is a graph showing the in vitro release profiles of PLL-insulin microparticles thermally treated at different temperatures.
  • Fig. 29A is a graph showing the serum insulin concentration versus time profiles in rats that have received a single subcutaneous injection of uncoated insulin microparticles, PLA-modified insulin microparticles treated at 28° C, and PLA-modified insulin microparticles treated at 4° C (Example 18); and
  • Fig. 29B is a graph showing the serum glucose depression concentration versus time profiles in rats that have received a single subcutaneous injection of uncoated insulin microparticles, PLA-modified insulin microparticles treated at 28° C, and PLA- modified insulin microparticles treated at 4° C (Example 18).
  • Active agent refers to naturally occurring, synthetic, or semi-synthetic materials (e.g., compounds, fermentates, extracts, cellular structures) capable of eliciting, directly or indirectly, one or more physical, chemical, and/or biological effects, in vitro and/or in vivo.
  • the active agent may be capable of preventing, alleviating, treating, and/or curing abnormal and/or pathological conditions of a living body, such as by destroying a parasitic organism, or by limiting the effect of a disease or abnormality by materially altering the physiology of the host or parasite.
  • the active agent may be capable of maintaining, increasing, decreasing, limiting, or destroying a physiologic body function.
  • the active agent may be capable of diagnosing a physiological condition or state by an in vitro and/or in vivo test.
  • the active agent may be capable of controlling or protecting an environment or living body by attracting, disabling, inhibiting, killing, modifying, repelling and/or retarding an animal or microorganism.
  • the active agent may be capable of otherwise treating (such as deodorizing, protecting, adorning, grooming) a body.
  • the active agent may further be referred to as a bioactive agent, a pharmaceutical agent (such as a prophylactic agent, a therapeutic agent), a diagnostic agent, a nutritional supplement, and/or a cosmetic agent, and includes, without limitation, prodrugs, affinity molecules, synthetic organic molecules, polymers, molecules with a molecular weight of 2kD or less (such as 1.5kD or less, or IkD or less), macromolecules (such as those having a molecular weight of 2kD or greater, preferably 5kD or greater), proteinaceous compounds, peptides, vitamins, steroids, steroid analogs, lipids, nucleic acids, carbohydrates, precursors thereof, and derivatives thereof.
  • Active agents may be ionic or non-ionic, may be neutral, positively charged, negatively charged, or zwitterionic, and may be used singly or in combination of two or more thereof. Active agents may be water insoluble, but more preferably are water soluble. Active agents may have an isoelectric point of 7.0 or greater, but preferably less than 7.0.
  • “Microparticle” refers to a particulate that is solid (including substantially solid or semi-solid, but excluding gel, liquid and gas), having an average geometric particle size (sometimes referred to as diameter) of less than 1 mm, preferably 200 microns or less, more preferably 100 microns or less, most preferably 10 microns or less.
  • the particle size may be 0.01 microns or greater, preferably 0.1 microns or greater, more preferably 0.5 microns or greater, and most preferably from 0.5 microns to 5 microns.
  • Average geometric particle size may be measured by dynamic light scattering methods (such as photocorrelation spectroscopy, laser diffraction, low-angle laser light scattering (LALLS), medium-angle laser light scattering (MALLS)), light obscuration methods (such as Coulter analysis method), or other methods (such as rheology, light or electron microscopy).
  • Particles for pulmonary delivery will have an aerodynamic particle size determined by time of flight measurements or Andersen Cascade Impactor measurements.
  • Microparticles may have a spherical shape (sometimes referred to as microspheres) and/or may be encapsulated (sometimes referred to as microcapsules). Certain microparticles may have one or more internal voids and/or cavities. Other microparticles may be free of such voids or cavities. Microparticles may be porous or, preferably non-porous. Microparticles may be formed from;, in part or in whole, one or more non-limiting materials, such as the active agents, carriers, polymers, stabilizing agents, and/or complexing agents disclosed herein.
  • Peptides refer to natural, synthetic, or semi-synthetic compounds formed at least in part from two or more of the same or different amino acids and/or imino acids.
  • Non-limiting examples of peptides include oligopeptides (such as those having less than 50 amino/imino acid monomer units, including dipeptides and tripeptides and the like), polypeptides, proteinaceous compounds as defined herein, as well as precursors and derivatives thereof (e.g., glycosylated, hyperglycosylated, PEGylated, FITC-labeled, salts thereof).
  • Peptides may be used singly or in combination of two or more thereof.
  • Peptides may be neutral, positively charged, negatively charged, or zwirterionic, and may be used singly or in combination of two or more thereof.
  • Proteinaceous compounds refer to natural, synthetic, semi-synthetic, or recombinant compounds of or related structurally and/or functionally to proteins, such as those containing or consisting essentially of ⁇ -amino acids covalently associated through peptide linkages.
  • Non-limiting proteinaceous compounds include globular proteins (e.g., albumins, globulins, histones), fibrous proteins (e.g., collagens, elastins, keratins), compound proteins (including those containing one or more non-peptide component, e.g., glycoproteins, nucleoproteins, mucoproteins, lipoproteins, metalloproteins), therapeutic proteins, fusion proteins, receptors, antigens (such as synthetic or recombinant antigens), viral surface proteins, hormones and hormone analogs, antibodies (such as monoclonal or polyclonal antibodies), enzymes, Fab fragments, cyclic peptides, linear peptides, and the like.
  • globular proteins e.g., albumins, globulins, histones
  • fibrous proteins e.g., collagens, elastins, keratins
  • compound proteins including those containing one or more non-peptide component, e.g
  • Non-limiting therapeutic proteins include bone morphogenic proteins, drug resistance proteins, toxoids, erythropoietins, proteins of the blood clotting cascade (e.g., Factor VII, Factor VIII, Factor IX, et al.), subtilisin, ovalbumin, alpha- 1 -antitrypsin (AAT), DNase, superoxide dismutase (SOD), lysozyme, ribonuclease, hyaluronidase, collagenase, human growth hormone (hGH), erythropoietin, insulin and insulin-like growth factors or their analogs, interferons, glatiramer, granulocyte-macrophage colony-stimulating factor, granulocyte colony-stimulating factor, desmopressin, leutinizing hormone release hormone (LHRH) agonists (e.g., leuprolide, goserelin, buserelin, gonadorelin, his
  • Nucleic acids refer to natural, synthetic, semi-synthetic, or recombinant compounds formed at least in part from two or more of the same or different nucleotides, and may be single-stranded or double-stranded.
  • Non-limiting examples of nucleic acids include oligonucleotides (such as those having 20 or less base pairs, e.g., sense, anti-sense, or missense), aptamers, polynucleotides (e.g., sense, anti-sense, or missense), DNA (e.g., sense, anti-sense, or missense), RNA (e.g., sense, anti-sense, or missense), siRNA, nucleotide acid constructs, single-stranded or double-stranded segments thereof, as well as precursors and derivatives thereof (e.g., glycosylated, hyperglycosylated, PEGylated, FITC-labeled, nucleosides, salts thereof). Nucleic acids may be neutral, positively charged, negatively charged, or zwitterionic, and may be used singly or in combination of two or more thereof.
  • Carbohydrates refer to natural, synthetic, or semi-synthetic compounds formed at least in part from monomeric sugar units.
  • Non-limiting carbohydrates include polysaccharides, sugars, starches, and celluloses, such as carboxymethylcellulose, dextrans, hetastarch, cyclodextrins, alginates, chitosans, chondroitins, heparins, as well as precursors and derivatives thereof (e.g., glycosylated, hyperglycosylated, PEGylated, FITC-labeled, salts thereof).
  • Carbohydrates may be ionic or non-ionic, may be neutral, positively charged, negatively charged, or zwitterionic, and may be used singly or in combination of two or more thereof.
  • Lipids refer to natural, synthetic, or semi-synthetic compounds that are generally amphiphilic.
  • the lipids typically comprise a hydrophilic component and a hydrophobic component.
  • Non-limiting examples include fatty acids, neutral fats, phosphatides, oils, glycolipids, surfactants, aliphatic alcohols, waxes, terpenes and steroids.
  • Lipids may be ionic or non-ionic, may be neutral, positively charged, negatively charged, or zwitterionic, and may be used singly or in combination of two or more thereof.
  • “Complexing agent” refers to a material capable of forming one or more non-covalent associations with the active agent.
  • the complexing agent is capable of facilitating the loading of one or more active agents into the microparticle, retaining the active agent(s) within the microparticle, and/or otherwise modifying the release of the active agent(s) from the microparticle.
  • Complexing agents may be ionic or non-ionic, may be neutral, positively charged, negatively charged, or zwitterionic, and may be used singly or in combination of two or more thereof.
  • Stabilizing used especially in conjunction with an agent (e.g., compound), a process, or a condition, refers to the capability of such agent, process or condition to, at least in part, form the microparticles (or a composition or formulation or kit containing such microparticles), facilitate the formation thereof, and/or enhance the stability thereof (e.g., the maintenance of a relatively balanced condition, like increased resistance against destruction, decomposition, degradation, and the like).
  • Non-limiting stabilizing processes or conditions include thermal input/output (e.g., heating, cooling), electromagnetic irradiation (e.g., gamma rays, X rays, UV, visible light, actinic, infrared, microwaves, radio waves), high-energy particle irradiation (e.g., electron beams, nuclear), and ultrasound irradiation.
  • thermal input/output e.g., heating, cooling
  • electromagnetic irradiation e.g., gamma rays, X rays, UV, visible light, actinic, infrared, microwaves, radio waves
  • high-energy particle irradiation e.g., electron beams, nuclear
  • ultrasound irradiation e.g., ultrasound irradiation.
  • Non-limiting stabilizing agents include lipids, proteins, polymers, carbohydrates, surfactants, salts (e.g., organic, inorganic, with cations that are monovalent or polyvalent, metallic, organic, or organometallic, and anions that are monovalent or polyvalent, organic, inorganic, or organometallic), as well as certain of the carriers, the active agents, the crosslinkers, the co-agents, and the complexing agents disclosed herein.
  • the stabilizing agents may be ionic or non-ionic, may be neutral, positively charged, negatively charged, or zwitterionic, and may be used singly or in combination of two or more thereof.
  • Micromolecule refers to a material capable of providing a three- dimensional (e.g., tertiary and/or quaternary) structure, and includes carriers and certain active agents of the present disclosure.
  • Non-limiting macromolecules used to form the microparticles include, inter alia, polymers, copolymers, proteins (e.g., enzymes, recombinant proteins, albumins like human serum albumin), peptides, lipids, carbohydrates, polysaccharides, nucleic acids, vectors (e.g., virus, viral particles), _
  • complexes and conjugates thereof e.g., by covalent and/or non-covalent associations, between two macromolecules like carbohydrate-protein conjugates, between an active agent and a macromolecule like hapten-protein conjugates, the active agent may or may not be capable of having a tertiary and/or quaternary structure), and mixtures of two or more thereof, preferably having a molecular weight of 1,500 or greater.
  • Macromolecules may be neutral, positively charged, negatively charged, or zwitterionic, and may be used singly or in combination of two or more thereof.
  • Spherical refers to a geometric shape that is at least “substantially spherical.” “Substantially spherical” means that the ratio of the longest length (i.e., one between two points on the perimeter and passes the geometric center of the shape) to the shortest length on any cross-section that passes through the geometric center is about 1.5 or less, preferably about 1.33 or less, more preferably 1.25 or less, spherical does not require a line of symmetry.
  • microparticles may have surface texturing (such as continuous or discrete lines, islands, lattice, indentations, channel openings, protuberances that are small in scale when compared to the overall size of the microparticles) and still be spherical. Surface contact there between is minimized in microparticles that are spherical, which minimizes the undesirable agglomeration of the microparticles. In comparison, microparticles that are crystals or flakes typically display significant agglomeration through ionic and/or non-ionic interactions at relatively large flat surfaces.
  • “Monodisperse size distribution” refers to a preferred microparticle size distribution in which the ratio of the volume diameter of the 90 th percentile (i.e., the average particle size of the largest 10% of the microparticles) to the volume diameter of the 10 th percentile (i.e., the average particle size of the smallest 10% of the microparticles) is about 5 or less, preferably about 3 or less, more preferably about 2 or less, most preferably about 1.5 to 1. Consequently, "polydisperse size distribution” refers to one where the diameter ratio described above is greater than 5, preferably greater than 8, more preferably greater than 10.
  • microparticles having a polydisperse size distribution may fill in the gaps between larger microparticles, thus possibly displaying large contact surfaces and significant agglomeration there between.
  • a Geometric Standard Deviation (GSD) of 2.5 or less, preferably 1.8 or less, may also be used to indicate a monodisperse size distribution. Calculation of GSD is known and understood to one skilled in the art.
  • Amorphous refers to materials and constructions that are “substantially amorphous,” such as microparticles having multiple non-crystalline domains (or lacking crystallinity altogether) or otherwise non-crystalline.
  • Substantially amorphous microparticles of the present disclosure are generally random solid particulates in which crystalline lattices constitute less than 50% by volume and/or weight of the microparticles, or are absent, and include semi-crystalline microparticles and non-crystalline microparticles as understood by one skilled in the art.
  • Solid refers to a state that includes at least substantially solid and/or semisolid, but excludes gel, liquid, and gas.
  • Preformed microparticle refers to a microparticle fabricated using one or more non-limiting methods, such as those known to one skilled in the art, without surface modification as described herein, having or capable of having on its outer surface a net surface electric charge that is positive, negative, or neutral.
  • a preformed microparticle is also referred to herein as "core microparticle” or “core.”
  • the preformed or core microparticle typically comprises one or more active agents and, optionally, one or more carriers, which, independently, may be compartmentalized in a portion of the preformed or core microparticle or preferably be distributed substantially homogeneously throughout the preformed microparticles.
  • the net surface charge preferably being non-zero, may be contributed primarily or at least, substantially, by the active agent(s) and/or the optional carrier(s).
  • Carrier refers to a compound, typically a macromolecule, having a primary function to provide a three-dimensional structure (including tertiary and/or quaternary structure).
  • the carrier may be unassociated or associated with the active agent (such as conjugates or complexes thereof) in forming microparticles as described above.
  • the carrier may further provide other functions, such as being an active agent, modify release profile of the active agent from the microparticle, and/or impart one or more particular properties to the microparticle (such as contribute at least in part to the net surface charge).
  • the carrier is a protein (such as albumins, preferably human serum albumin) having a molecular weight of 1500 Daltons or greater.
  • Polymer or “polymeric” refers to a natural, synthetic, or semi-synthetic molecule having in a main chain or ring structure two or more repeating monomer units. Polymers broadly include dimers, trimers, tetramers, oligomers, higher molecular weight polymer, adducts, homopolymers, random copolymers, pseudo-copolymers, statistical copolymers, alternating copolymers, periodic copolymer, bipolymers, terpolymers, quaterpolymers, other forms of copolymers, substituted derivatives thereof, and mixtures thereof, and narrowly refer to molecules having 10 or more repeating monomer units.
  • Polymers may be linear, branched, block, graft, monodisperse, polydisperse, regular, irregular, tactic, isotactic, syndiotactic, stereoregular, atactic, stereoblock, single-strand, double-strand, star, comb, dendritic, and/or ionomeric, may be ionic or non-ionic, may be neutral, positively charged, negatively charged, or zwitterionic, and may be used singly or in combination of two or more thereof.
  • suspension refers to a mixture, preferably finely divided, of two or more phases (e.g., solid, liquid, gas), such as solid in liquid, liquid in liquid, gas in liquid, solid in solid, solid in gas, liquid in gas, and the like.
  • the suspension or dispersion may preferably remain stable for extended periods of time (e.g., minutes, hours, days, weeks, months, years).
  • Resuspending refers to changing microparticles from a non-flowable (e.g., solid) state to a flowable (e.g., liquid) state by adding a flowable medium (e.g., a liquid), while retaining most or all of the characteristics of the microparticles.
  • the liquid may be, for example, aqueous, aqueous miscible, or organic.
  • Charged and electrically charged refer interchangeably to the capability of providing one, two, three, or more formal units of electrical charges of the same or opposite sign and/or the presence of such charges (i.e., “charged” refers to chargeable and/or charged).
  • the electrical charges may be provided by and/or be present in the form of one or more of the same or different organic and/or organometallic moieties (e.g., ionic groups, ionizable groups, precursors thereof) in the compound (e.g., polyelectrolytes, proteins) or the structure (e.g., the preformed microparticle, the monolayers) of interest, preferably when subjected to certain conditions (such as in solution or suspension).
  • organic and/or organometallic moieties e.g., ionic groups, ionizable groups, precursors thereof
  • the compound e.g., polyelectrolytes, proteins
  • the structure e.g., the preformed microparticle, the monolayers
  • Charged compound and “electrically charged compound” refer interchangeably to a single compound that is charged as described above, or a combination of two or more different compounds in unassociated and/or associated forms (e.g., conjugates, aggregates, and/or complexes thereof), each of which independently has and/or is capable of having a net charge of the same sign.
  • “Monolayer” refers to a single layer formed over a three-dimensional substrate, from a composition of one or more compounds (such as a charged compound as described above).
  • the monolayer may be a continuous and nonporous monolayer, a continuous and porous monolayer (such as a lattice network), a non-continuous monolayer of a plurality of discrete elements (e.g., islands, strips, clusters, etc.), or a combination thereof.
  • the monolayer will be decomposable or degradable, such as biodegradable, enzymatically or hydrolytically degradable and the like, to allow for non- diffusional release of an active agent (from the microparticle) over which the monolayer is deposited.
  • the monolayer may have a thickness of 100 nm or less, preferably 50 rnn or less, more preferably 20 nm or less, most preferably 10 nm or less.
  • the monolayer is formed through self-assembly of a charged compound.
  • "Saturated monolayer” refers to a monolayer as defined above that is incapable of further incorporating, cumulatively, an excess amount of the composition forming the monolayer when subjected to the same set of conditions under which the monolayer is formed. Saturated monolayers are preferred monolayers used in surface- modified microparticles.
  • Net charge and "net electric charge” are used interchangeably and refer to the sum of all formal units of electric charge a charged compound is capable of having or has, such as in a flowable medium under certain conditions (preferably in a solution of certain pH).
  • the net charge may be positive, negative, or zero (such as in zwitterionic compounds), and is condition-dependent (e.g., solvent, pH).
  • Network surface charge and “net surface electric charge” are used interchangeably and refer to an overall cumulative electric charge on an outermost surface of a three-dimensional structure (e.g., a microparticle, a monolayer). The net surface charge may be positive, negative, or zero, and is condition-dependent (e.g., solvent, pH).
  • Ambient temperature refers to a temperature of around room temperature, typically in a range of about 20 0 C to about 40°C.
  • Subject or “patient” refers to animals, including vertebrates like mammals, preferably humans.
  • Regular of a subject refers to a localized internal or external area or portion of the subject (e.g., an organ), or a collection of areas or portions throughout the entire subject (e.g., lymphocytes).
  • Non-limiting examples of such regions include pulmonary region (e.g., lung, alveoli, gastrointestinal region (e.g., regions defined by esophagus, stomach, small large intestines, and rectum), cardiovascular region (e.g., myocardial tissue), renal region (e.g., the region defined by the kidney, the abdominal aorta, and vasculature leading directly to and from the kidney), vasculature (i.e., blood vessels, e.g., arteries, veins, capillaries, and the like), circulatory system, healthy or diseased tissues, benign or malignant (e.g., tumorous or cancerous) tissues, lymphocytes, receptors, organs and the like, as well as regions to be imaged with diagnostic imaging, regions to be administered and/or treated with an active agent, regions to be targeted for the delivery of an active agent, and regions of elevated temperature.
  • pulmonary region e.g., lung, alveoli
  • gastrointestinal region e.g., regions defined
  • Therapeutic refers to any pharmaceutic, drug, prophylactic agent, contrast agent, or dye useful in the treatment (including prevention, diagnosis, alleviation, suppression, remission, or cure) of a malady, affliction, disease or injury in a subject.
  • Therapeutically useful peptides and nucleic acids may be included within the meaning of the term “pharmaceutic” or "drug.”
  • Affinity molecule refers to any material or substance capable of promoting binding and/or targeting of regions in vivo and/or tissues/receptors in vitro.
  • Affinity molecules including receptors and targeting ligands, may be natural, synthetic, or semisynthetic, may be ionic or non-ionic, may be neutral, positively charged, negatively charged, or zwitterionic, and may be used singly or in combination of two or more thereof.
  • Non-limiting affinity molecules include proteinaceous compounds (e.g., antibodies, antibody fragments, hormones, hormone analogues, glycoproteins and lectins), peptides, polypeptides, amino acids, sugars, saccharides (e.g., monosaccharides, polysaccharides, carbohydrates), vitamins, steroids, steroid analogs, cofactors, active agents, nucleic acids, viruses, bacteria, toxins, antigens, other ligands, precursors thereof, and derivatives thereof.
  • proteinaceous compounds e.g., antibodies, antibody fragments, hormones, hormone analogues, glycoproteins and lectins
  • peptides e.g., polypeptides, amino acids, sugars, saccharides (e.g., monosaccharides, polysaccharides, carbohydrates), vitamins, steroids, steroid analogs, cofactors, active agents, nucleic acids, viruses, bacteria, toxins, antigens, other ligands, precursors thereof, and derivative
  • Precursor refers to any material or substance capable of being converted to a desired material or substance, preferably through a chemical and/or biochemical reaction or pathway, such as anchoring a precursor to a material.
  • Non-limiting precursor moieties include maleimide groups, disulfide groups (e.g., ortho-pyridyl disulfide), vinylsulfone groups, azide groups, and ⁇ -iodo acetyl groups.
  • Derivative refers to any material or substance formed from a parent material or substance, preferably through a chemical and/or biochemical reaction or pathway considered routine by one of ordinary skill in the art.
  • Non-limiting examples of derivatives include glycosylated, hyperglycosylated, PEGylated, FITC-labelled, protected with protecting groups (e.g., benzyl for alcohol or thiol, t-butoxycarbonyl for amine), as well as salts, esters, amides, conjugates, complexes, manufacturing related compounds, and metabolites thereof.
  • Salts may be organic or inorganic, with cations that are monovalent or polyvalent, metallic, organic, or organometallic, and anions that are monovalent or polyvalent, organic, inorganic, or organometallic.
  • Preferred salts are pharmaceutically acceptable, and include, without limitation, mineral or organic acid salts of basic residues (e.g., amines), alkali or organic salts of acidic residues (e.g., carboxylic acids), and the like, such as conventional non-toxic salts or the quaternary ammonium salts of the parent compound formed from non-toxic inorganic acids (e.g., hydrochloric, hydrobromic, sulfuric, sulfamic, phosphoric, nitric) and organic acids (e.g., acetic, propionic, succinic, glycolic, stearic, lactic, malic, tartaric, citric, ascorbic, pamoic, maleic, hydroxymaleic, phenylacetic, glutamic, be
  • Analog refers to a compound having a chemically modified form of a principle compound or class thereof, which maintains the pharmaceutical and/or pharmacological activities characteristic of the principle compound or class.
  • Prodrug refers to any covalently bonded carriers that release an active agent in vivo when administered to a subject. Prodrugs are known to enhance numerous desirable qualities (e.g., solubility, bioavailability, manufacturing) of the active agents.
  • Prodrugs may be prepared by modifying functional groups (e.g., hydroxy, amino, carboxyl, and/or sulfhydryl groups) present in the active agent in such a way that the modifications are reversed (e.g., modifier group cleaved), either in routine manipulation or in vivo, to afford the original active agent.
  • the transformation in vivo may be, for example, as the result of some metabolic process, such as chemical or enzymatic hydrolysis of a carboxylic, phosphoric or sulphate ester, or reduction or oxidation of a susceptible functionality.
  • "Metabolite” refers to a form of a compound obtained in a subject body by action of the body on the administered form of the compound. For example, a demethylated metabolite may be obtained in the body after administration of a methylated compound bearing a methyl group. Metabolites may themselves have biological, preferably therapeutic, activities.
  • Diagnostic agent refers to any material or substance useful in connection with methods for perceptually observing (e.g., imaging) a normal or abnormal biological condition or state, or detecting the presence or absence of a pathogen or a pathological condition.
  • Non-limiting diagnostic agents include contrast agents and dyes for use in connection with radiography imaging (e.g., X-ray imaging), ultrasound imaging, magnetic resonance imaging, computed tomography, positron emission tomography imaging, and the like. Diagnostic agents further include any other agents useful in facilitating diagnosis in vivo and/or in vitro, whether or not imaging methodology is employed.
  • Cross-link generally refer to the linking of two or more materials and/or substances, including any of those disclosed herein, through one or more covalent and/or non-covalent (e.g., ionic) associations.
  • Cross- linking may be effected naturally (e.g., disulfide bonds of cystine residues) or through synthetic or semi-synthetic routes, for example, optionally in the presence of one or more cross-linkers (i.e., a molecule X by itself capable of reacting with two or more materials/substances Y and Z to form a cross-link product Y-X-Z, where the associations of Y-X and X-Z are independently covalent and/or non-covalent), initiators (i.e., a molecule by itself capable of providing reactive species like free radicals for the cross-link reaction, e.g., thermally decomposable initiators like organic peroxides, azo initiators, and carbon-carbon initiators, actinically decomposable initiators like photoinitiators of various wavelengths), activators (i.e., a molecule A capable of reacting with a first material/substance Y to form an activated intermediate [A-Y], which in turn reacts with a
  • Covalent association refers to an intermolecular interaction (e.g., a bond) between two or more individual molecules that involves the sharing of electrons in the bonding orbitals of two atoms.
  • Non-covalent association refers to an intermolecular interaction between two or more individual molecules without involving a covalent bond. Intermolecular interaction depends on, for example, polarity, electric charge, and/or other characteristics of the individual molecules, and includes, without limitation, electrostatic (e.g., ionic) interactions, dipole-dipole interactions, van der Waal's forces, and combinations of two or more thereof.
  • electrostatic e.g., ionic
  • Electrostatic interaction refers to an intermolecular interaction between two or more positively or negatively charged moieties/groups, which may be attractive when two are oppositely charged (i.e., one positive, another negative), repulsive when two charges are of the same sign (i.e., two positive or two negative), or a combination thereof.
  • Dipole-dipole interaction refers an intermolecular attraction between two or more polar molecules, such as a first molecule having an uncharged, partial positive end ⁇ + (e.g., electropositive head group like the choline head group of phosphatidylcholine) and a second molecule having an uncharged, partial negative end ⁇ " (e.g., an electronegative atom like heteroatom O, N, or S in a polysaccharide).
  • a first molecule having an uncharged, partial positive end ⁇ + e.g., electropositive head group like the choline head group of phosphatidylcholine
  • a second molecule having an uncharged, partial negative end ⁇ " e.g., an electronegative atom like heteroatom O, N, or S in a polysaccharide
  • Dipole-dipole interaction also refers to intermolecular hydrogen bonding in which a hydrogen atom serves as a bridge between electronegative atoms on separate molecules and in which a hydrogen atom is held to a first molecule by a covalent bond and to a second molecule by electrostatic forces.
  • Hydrogen bond refers to an attractive force or bridge between a hydrogen atom covalently bonded to a first electronegative atom (e.g., O, N, S) and a second electronegative atom, where the first and second electronegative atoms may be in two different molecules (intermolecular hydrogen bonding) or in a single molecule (intramolecular hydrogen bonding).
  • a first electronegative atom e.g., O, N, S
  • second electronegative atoms may be in two different molecules (intermolecular hydrogen bonding) or in a single molecule (intramolecular hydrogen bonding).
  • Van der Waal's forces refers to the attractive forces between non-polar molecules that are accounted for by quantum mechanics. Van der Waal's forces are generally associated with momentary dipole moments induced by neighboring molecules undergoing changes in electron distribution.
  • Hydrophilic interaction refers to an attraction toward water molecules, where a material/compound or a portion thereof may bind with, absorb, and/or dissolve in water. This may result in swelling and/or the formation of reversible hydrogels.
  • Hydrophilic interaction refers to a repulsion against water molecules, where a material/compound or a portion thereof do not bind with, absorb, or dissolve in water.
  • Biocompatible refers to materials/substances that are generally not injurious to biological functions and do not result in unacceptable toxicity (e.g., allergenic responses or disease states).
  • association with and “associated with” refer in general to the one or more interactions between, and/or incorporation of, different materials (typically those that are part of the microparticles), one or more of such materials and one or more structures (or portions thereof) of the microparticles, and different structures (or portions thereof) of the microparticles.
  • the materials of the microparticles include, without limitation, ions such as monovalent and polyvalent ions disclosed herein, as well as compounds such as active agents, stabilizing agents, cross-link agents, charged or uncharged compounds, the various polymers disclosed herein, and combinations of two or more thereof.
  • the structures of the microparticles and portions thereof include, without limitation, core, core microparticle, preformed microparticle, monolayer, intermediate microparticle, surface- modified microparticle, portions of such structures (such as outer surfaces, inner surfaces), domains between such structures and portions thereof, and combinations of two or more thereof.
  • Various associations, being reversible or irreversible, migratory or non- migratory, may be present singly or in combination of two or more thereof.
  • Non-limiting associations include, without limitation, covalent and/or non-covalent associations (e.g., covalent bonding, ionic interactions, electrostatic interactions, dipole-dipole interactions, hydrogen bonding, van der Waal's forces, cross-linking, and/or any other interactions), encapsulation in layer/membrane, compartmentalization in center or vesicles or between two layers/membranes, homogeneous integration throughout the microparticle or in a portion thereof (e.g., containment in, adhesion to, and/or affixation to center or layer or vesicle or an inner and/or outer surface thereof; interspersion, conjugations, and/or complexation between different materials).
  • covalent and/or non-covalent associations e.g., covalent bonding, ionic interactions, electrostatic interactions, dipole-dipole interactions, hydrogen bonding, van der Waal's forces, cross-linking, and/or any other interactions
  • encapsulation in layer/membrane
  • tissue refers generally to an individual cell or a plurality or aggregate of cells specialized and capable of performing one or more particular functions.
  • tissue examples include membranous tissues, (e.g., endothelium, epithelium), blood, laminae, connective tissue (e.g., interstitial tissue), organs (e.g., myocardial tissue, myocardial cells, cardiomyocites), abnormal cell(s) (e.g., tumors).
  • Receptor refers to a molecular structure within a cell or on its surface, generally characterized by its selective binding of a specific substance, e.g., ligand.
  • Non- limiting receptors include cell-surface receptors for peptide hormones, neurotransmitters, antigens, complement fragments, and immunoglobulins, and cytoplasmic receptors for steroid hormones.
  • Controlled release refers to a predetermined in vivo and/or in vitro release (e.g., dissolution) profile of an active agent, as compared to the release profile of the active agent in its native form.
  • the active agent is preferably associated with a microparticle or a composition or formulation containing such a microparticle, as disclosed herein, such that one or more aspects of its release kinetics (e.g., initial burst, quantity and/or rate over a specified time period or phase, cumulative quantity over a specific time period, length of time for total release, pattern and/or profile, etc.) are increased, decreased, shortened, prolonged, and/or otherwise modified as desired.
  • Non-limiting examples of controlled release include immediate/instant release (i.e., initial burst or rapid release), extended release, sustained release, prolonged release, delayed release, modified release, and/or targeted release, occurring individually, in combination of two or more thereof, or in the absence of one or more thereof (e. g. extended or sustained release in the absence of an initial burst).
  • Extended release refers to the release of an active agent, preferably in association with a microparticle or a composition or formulation containing such a microparticle, as disclosed herein, over a time period longer than the free aqueous diffusion period of the active agent in its native form.
  • the extended release period may be hours (e.g., at least about 1, 2, 5, or 10 hours), days (e.g., at least about 1, 2, 3, 4, 5, 6, 7, 8, 10, 15, 20, 30, 40, 45, 60, or 90 days), weeks (at least about 1, 2, 3, 4, 5, 6, 10, 15, 20, 30, 40, or 50 weeks), months (at least about 1, 2, 3, 4, 6, 9, or 12 months), about 1 or more years, or a range between any two of the time periods.
  • the pattern of an extended release may be continuous, periodic, sporadic, or a combination thereof.
  • sustained release refers to an extended release of an active agent such that a functionally significant level of the active agent (i.e., a level capable of bring about the desired function of the active agent) is present at any time point of the extended release period, preferably with a continuous and/or uniform release pattern.
  • sustained release profiles include those, when displayed in a plot of release time (x-axis) versus cumulative release (y-axis), showing at least one upward segment that is linear, step-wise, zig-zagging, curved, and/or wavy, over a time period of 1 hour or longer.
  • compositions “formed from” and “formed of a list of recited components be a composition comprising at least these recited components, and can further include other non-recited components during formulation of the composition.
  • Examples provided herein, including those following "such as” and “e.g.,” are considered as illustrative only of various aspects of the present disclosure and embodiments thereof, without being specifically limited thereto.
  • each of the surface-modified microparticles of the present disclosure preferably contains an amorphous (e.g., such as free of crystalline structures) and solid preformed microparticle associated, at least at its outer surface, with at least one monolayer containing at least one charged compound.
  • the preformed microparticle contains at least one active agent and/or at least one macromolecule having a molecular weight of 4,500 Daltons or greater.
  • the macromolecule may be the active agent or may be different from the active agent.
  • the macromolecule may be a carrier, a stabilizing agent, or a complexing agent (e.g., proteinaceous compounds, polyelectrolytes).
  • the active agent and/or the macromolecule may constitute 40% to 100% or less, and typically at least 80%, such as 90% or more or 95% or more, by weight of the preformed microparticle.
  • the active agent and/or the macromolecule is/are distributed homogeneously throughout the core microparticle.
  • An outer surface of the preformed microparticle carries a net surface charge, which may be attributed, at least in part, and more typically in large part, to the active agent and/or the macromolecule, especially when the outer surface is formed of the active agent and/or the macromolecule.
  • the preformed microparticle may be free of covalent crosslmking, hydrogel, lipids, and/or encapsulation.
  • the preformed microparticle may contain one or more charged compounds, covalent crosslinking, and/or encapsulation.
  • the one or more charged compounds in the preformed microparticle may be distributed homogeneously throughout the preformed microparticle, or compartmentalized in specific portions thereof, such as in a layer.
  • the preformed microparticle may preferably have a particle size of 10 ⁇ m or less, and may have a monodisperse or polydisperse size distribution.
  • Methods of pre-forming the preformed microparticle are not particularly limiting, and include those disclosed in U.S. Patent No. 6,458,387 and U.S. Patent Publication No. 2005/0142206, which are incorporated herein by reference in their entirety.
  • a single flowable continuous phase system (such as liquid, gas, or plasma, preferably a solution or suspension) is formulated to contain one or more active agents, a medium, and one or more phase-separation enhancing agents (PSEAs).
  • PSEAs phase-separation enhancing agents
  • the medium is preferably a liquid solvent (e.g., hydrophilic or hydrophobic organic solvents, water, buffers, aqueous-miscible organic solvents, and combinations of two or more thereof), more preferably an aqueous or aqueous-miscible solvent.
  • Suitable organic solvents include, without limitation, methylene chloride, chloroform, acetonitrile, ethylacetate, methanol, ethanol, pentane, the likes thereof, and combinations of two or more thereof (such as a 1:1 mixture of methylene chloride and acetone).
  • the active agent and the PSEA may independently be dissolved, suspended, or otherwise homogeneously distributed within the medium.
  • the active agent When subjecting the flowable system to certain conditions (such as a temperature below the phase transition temperature of the active agent in the medium), the active agent undergoes a liquid-solid phase separation and forms a discontinuous, preferably solid, phase (such as a plurality of core microparticles suspended in the medium), while the PSEA remains in the continuous phase (such as being dissolved in the medium).
  • certain conditions such as a temperature below the phase transition temperature of the active agent in the medium
  • the active agent undergoes a liquid-solid phase separation and forms a discontinuous, preferably solid, phase (such as a plurality of core microparticles suspended in the medium), while the PSEA remains in the continuous phase (such as being dissolved in the medium).
  • the medium can be organic, containing an organic solvent or a mixture of two or more inter-miscible organic solvents, which may independently be aqueous- miscible or aqueous-immiscible.
  • the solution can also be an aqueous-based solution containing an aqueous medium or an aqueous-miscible organic solvent or a mixture of aqueous-miscible organic solvents or combinations thereof.
  • the aqueous medium can be water, a buffer (e.g., normal saline, buffered solutions, buffered saline), and the like.
  • Suitable aqueous-miscible organic solvents may be monomers or polymers, and include, but are not limited to, N-methyl-2-pyrrolidinone (N-methyl-2-pyrrolidone), 2- pyrrolidinone (2-pyrrolidone), l,3-dimethyl-2-imidazolidinone (DMI) 5 dimethylsulfoxide, dimethylacetamide, acetic acid, lactic acid, acetone, methyl ethyl ketone, acetonitrile, methanol, ethanol, n-propanol, isopropanol, 3-pentanol, benzyl alcohol, glycerol, tetrahydrofuran (THF), polyethylene glycol (PEG, e.g., PEG-4, PEG-8, PEG-9, PEG- 12, PEG-14, PEG-16, PEG-120, PEG-75, PEG-150), PEG esters (e.g., PEG-4 d
  • a solution of the PSEA in a first solvent in which the PSEA is soluble in or miscible with the first solvent.
  • the active agent is mixed in, directly or as a second solution in a second solvent, with the first solution.
  • the first and second solvent may be the same or at least miscible with each other.
  • the active agent is added at a temperature equal to or lower than ambient temperature, particularly when the active agent is a heat labile molecule such as certain proteinaceous compounds.
  • the system may be heated to increase solubility of the active agent in the system, as long as the activity of the active agent is not adversely affected.
  • the PSEA while remaining in the liquid continuous phase, enhances and/or induces a liquid-solid phase separation of the active agent from the solution (such as by reducing solubility of the active agent), forming the core microparticles (the solid discontinuous phase), which may preferably be microspheres.
  • Suitable PSEA compounds include, but are not limited to, natural and synthetic polymers, linear polymers, branched polymers, cyclo-polymers, copolymers (random, block, grafted, such as poloxamers, particularly PLURONIC® Fl 27 and F68), terpolymers, amphiphilic polymers, carbohydrate-based polymers, polyaliphatic alcohols, poly(vinyl)polymers, polyacrylic acids, polyorganic acids, polyamino acids, polyethers, polyesters, polyimides, polyaldehydes, polyvinylpyrrolidone (PVP), and surfactants.
  • natural and synthetic polymers linear polymers, branched polymers, cyclo-polymers, copolymers (random, block, grafted, such as poloxamers, particularly PLURONIC® Fl 27 and F68), terpolymers, amphiphilic polymers, carbohydrate-based polymers, polyaliphatic alcohols, poly(vinyl)poly
  • Suitable or exemplary PSEAs include, without limitation, polymers acceptable as pharmaceutical additives, such as PEGs (e.g., PEG 200, PEG 300, PEG 3350, PEG 800O 5 PEG 10000, PEG 20000, etc.), poloxamers, PVP, hydroxyethylstarch, amphiphilic polymers, as well as non-polymers (such as mixtures of propylene glycol and ethanol).
  • Conditions capable of enhancing, inducing, promoting, controlling, suppressing, retarding, or otherwise affecting the liquid-solid phase separation include, without limitation, changes in temperature, pressure, pH, ionic strength and/or osmolality of the solutions, concentrations of the active agent and/or the PSEA, the likes thereof, as well as rates of such changes, and combinations of two or more thereof. Such conditions may desirably be applied before and up to the phase separation, or even during the phase separation. In one example, the system is exposed to a temperature below the phase transition temperature of the active agent therein, alone or in combination with adjustments to the concentrations of the active agent and/or the PSEA, as described in U.S.
  • FPDAs Freezing point depressing agents
  • FPDAs Freezing point depressing agents
  • Suitable FPDAs include, without limitation, propylene glycol, sucrose, ethylene glycol, alcohols (e.g., ethanol, methanol), and aqueous mixtures thereof.
  • the preformed microparticles may further comprise one or more excipients that negligibly affect the phase separation.
  • the excipient may imbue the core microparticles and/or the compounds therein (e.g., the active agent, the optional carrier) with additional characteristics such as increased stability, controlled release of the active agent from the preformed microparticles, and/or modified permeation of the active agent through biological tissues.
  • Suitable excipients include, but are not limited to, carbohydrates (e.g., trehalose, sucrose, mannitol), polyvalent cations (preferably metal cations, e.g., Zn 2+ , Mg 2+ , Ca 2+ , Cu 2+ , Fe 2+ , Fe 3+ ), anions (e.g., CO 3 2" , SO 4 2" ), amino acids (such as glycine), lipids, phospholipids, fatty acids and esters thereof, surfactants, triglycerides, bile acids and conjugates and salts thereof (e.g., cholic acid, deoxycholic acid, glycocholate, taurocholate, sodium cholate), and any polymers disclosed herein.
  • carbohydrates e.g., trehalose, sucrose, mannitol
  • polyvalent cations preferably metal cations, e.g., Zn 2+ , Mg 2+ , Ca 2+ , Cu 2
  • the preformed microparticles may optionally be separated from the solution and washed prior to the surface modification as disclosed herein, or be surface-modified without separation or washing. Separation means include, without limitation, centrifugation, dialysis, sedimentation (creaming), phase separation, chromatography, electrophoresis, precipitation, extraction, affinity binding, filtration, and diafiltration.
  • the washing medium may be aqueous, o r
  • SRAs solubility reducing agents
  • Preferred SRAs are capable of forming insoluble complexes with the active agents and/or carriers in the microparticles, and include, without limitation, compounds such as salts that comprise divalent or polyvalent cations (such as those disclosed herein).
  • the washing medium may be organic, or aqueous but containing at least one SRA or precipitating agent (such as ammonium sulfate).
  • the washing medium is the same solution used in the phase separation reaction, such as an aqueous solution including approximately 16% (w/v) PEG and 0.7% (w/v) NaCl.
  • the washing medium has a low boiling point for easy removal by, for example, lyophilization, evaporation, or drying.
  • the washing medium may be a supercritical fluid or a fluid near its supercritical point, used alone or in combination with a co-solvent.
  • Supercritical fluids may be solvents for the PSEAs, but not for the preformed microparticles.
  • Non-limiting examples of supercritical fluids include liquid CO 2 , ethane, and xenon.
  • Non-limiting examples of co-solvents include acetonitrile, dichloromethane, ethanol, methanol, water, and 2-propanol.
  • active agents with varying degrees of solubility in water may be employed in the microparticles described herein. While water insoluble active agents may be used, water soluble active agents are preferred.
  • the active agent may be a pharmaceutical agent.
  • the pharmaceutical agent includes, without limitation, adjuvants, adrenergic agents, adrenergic blocking agents, adrenocorticoids, adrenolytics, adrenomimetics, alkaloids, alkylating agents, allosteric inhibitors, anabolic steroids, analeptics, analgesics, anesthetics, anorexiants, antacids, anthelmintics, anti-allergic agents, antiangiogenesis agents, anti-arrhythmic agents, anti-bacterial agents, antibiotics, antibodies, anticancer agents, anticholinergic agents, anticholinesterases, anticoagulants, anticonvulsants, antidementia agents, antidepressants, antidiabetic agents, antidiarrheals, antidotes, antiepileptics, antifolics, antifungals, antigens, antihelmintics, antihistamine
  • the active agent may be used individually or in combinations of two or more thereof.
  • the active agent is a prophylactic and/or therapeutic agent that includes, but is not limited to, peptides, carbohydrates, nucleic acids, other compounds, precursors and derivatives thereof, and combinations of two or more thereof.
  • the active agent may be a cosmetic agent.
  • Non-limiting cosmetic agents include inter-alia emollients, humectants, free radical inhibitors, antiinflammatories, vitamins, depigmenting agents, anti-acne agents, antiseborrhoeics, keratolytics, slimming agents, skin coloring agents and sunscreen agents.
  • Non-limiting compounds useful as cosmetic agents include linoleic acid, retinol, retinoic acid, ascorbic acid alkyl esters, polyunsaturated fatty acids, nicotinic esters, tocopherol nicotinate, unsaponifiables of rice, soybean or shea, ceramides, hydroxy acids such as glycolic acid, selenium derivatives, antioxidants, beta-carotene, gamma-orizanol and stearyl glycerate.
  • the cosmetic agents may be commercially available and/or prepared by known techniques.
  • the active agent may be a nutritional supplement.
  • nutritional supplements include proteins, carbohydrates, water-soluble vitamins (e.g., vitamin C, B-complex vitamins, and the like), fat-soluble vitamins (e.g., vitamins A, D, E 5 K, and the like), and herbal extracts.
  • the nutritional supplements may be commercially available and/or prepared by known techniques.
  • the active agent may be a compound having a molecular weight of 2kD or less.
  • Non-limiting examples of such compounds include steroids, beta- agonists, anti-microbials, antifungals, taxanes (antimitotic and antimicrotubule agents), amino acids, aliphatic compounds, aromatic compounds, and urea compounds.
  • the active agent may be a therapeutic agent for prevention and/or treatment of pulmonary disorders.
  • Non-limiting examples of such agents include steroids, beta-agonists, anti-fungals, anti-microbial compounds, bronchial dialators, antiasthmatic agents, non-steroidal anti-inflammatory agents (NSAIDS), AAT, and agents to treat cystic fibrosis.
  • NSAIDS non-steroidal anti-inflammatory agents
  • Non-limiting examples of steroids include beclomethasone (such as beclomethasone dipropionate), fluticasone (such as fluticasone propionate), budesonide, estradiol, fludrocortisone, flucinonide, triamcinolone (such as triamcinolone acetonide), fmnisolide, and salts thereof.
  • Non-limiting examples of beta-agonists include salmeterol xinafoate, formoterol fumarate, levo-albuterol, bambuterol, tulobuterol, and salts thereof.
  • Non-limiting examples of anti-fungal agents include itraconazole, fluconazole, amphotericin B, and salts thereof.
  • the active agent may be a diagnostic agent.
  • diagnostic agents include x-ray imaging agents and contrast media.
  • x-ray imaging agents include ethyl 3,5-diacetamido-2,4,6-triiodobenzoate (WTN-8883, ethyl ester of diatrazoic acid); 6-ethoxy-6-oxohexyl-3,5-bis(acetamido)-2,4,6- triiodobenzoate (WIN 67722); ethyl-2-(3,5-bis(acetamido)-2,4,6-triiodobenzoyloxy)- butyrate (WIN 16318); ethyl diatrizoxyacetate (WIN 12901); ethyl 2-(3,5-bis(acetamido> 2,4,6-triiodobenzoyloxy) ⁇ ro ⁇ ionate (WIN 16923); N-ethyl 2-(3,5-bis(acetamido> 2,4,
  • contrast agents desirably disintegrate relatively rapidly under physiological conditions, thus minimizing any particle associated inflammatory response. Disintegration may result from enzymatic hydrolysis, solubilization of carboxylic acids at physiological pH, or other mechanisms.
  • poorly soluble i ⁇ dinated carboxylic acids such as iodipamide, diatrizoic acid, and metrizoic acid, along with hydrolytically labile iodinated species such as WIN 67721, WIN 12901, WIN 68165, and WIN 68209 or others may be preferred.
  • the active agents may be used in a combination of two or more thereof.
  • Non-limiting examples include a steroid and a beta-agonist, e.g., fluticasone propionate and salmeterol, budesonide and formeterol, etc.
  • Preformed microparticles may be substantially free of internal voids and/or cavities (such as being free of vesicles), substantially free of encapsulation, substantially free of lipids, substantially free of hydrogel or swelling, substantially and non-porous, amorphous solid, and/or spherical as those terms are defined herein.
  • Preformed microparticles may have multiple surface channel openings, the diameter of which are generally 100 nm or less, preferably 10 nm or less, more preferably 5 nm or less, most preferably 1 nm or less.
  • Preformed microparticles may have an overall density of 0.5 g/cm or greater, preferably 0.75 g/cm or greater, more preferably 0.85 g/cm or greater. The density may be generally up to about 2 g/cm 3 , preferably 1.75 g/cm 3 or less, more preferably 1.5 g/cm 3 or less.
  • Preformed microparticles may exhibit a high payload of the at least one active agent. Depending on the formulation and the physical/chemical nature of the compounds, there are typically at least 1000 or more, such as a few million to hundreds of millions of the active agent molecules in each of the preformed microparticles.
  • the weight percentage of the active agent in the preformed microparticle may be any of the amount below or greater, or any ranges there between, but less than 100%: 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 98%, 99%.
  • Surface modification of the preformed microparticles is achieved, without limitation, by forming, in a controlled manner, at least one monolayer containing at least one charged compound about the preformed microparticle.
  • each monolayer containing at least one charged compound contains different charged compounds, and preferably each carries on its outer surface a net surface charge that is different in sign and/or value from that of the preceding one and/or the subsequent one, if present.
  • Deposition of such monolayers one at a time allows for optimal control over various properties of the resulting microparticles, allowing one to tailor or "fine-tune" the microparticles to achieve a desired result.
  • the monolayer immediately about the performed microparticle contains one or more charged compounds, each independently having a net charge that is opposite in sign to the net surface charge of the core microparticle.
  • the preformed microparticle may at least, in part, be penetrable by the charged compound in the formed monolayer.
  • An outer surface of the formed monolayer may carry a net surface charge that is different from, preferably opposite in sign to, that of the preformed microparticle outer surface, especially when the formed monolayer is a saturated monolayer as defined herein.
  • the charged compounds may include one or more of polyelectrolytes, charged polyaminoacids, charged polysaccharides, polyionic polymers, charged proteinaceous compounds, charged peptides, charged lipids optionally in combination with uncharged lipids, charged lipid structures, and derivatives thereof.
  • the surface-modified microparticle may further contain one or more additional alternatingly charged monolayers, such that the surface-modified microparticle has a desired release profile of the active agent. This number is not particularly limited, but may typically be between 1 to 7, such as 2, 3, 4, 5, or 6.
  • one or more of such charged monolayers may independently have one or more of the same or different active agents, such as an affinity molecule, especially a targeting ligand, associated covalently and/or non-covalently thereto, preferably on their respective outer surfaces.
  • the core microparticle may have one or more portions, such as a center or an underlying layer (a charged monolayer, for example), containing at least one such active agent, preferably on the outer surface of the portion.
  • the preformed microparticle, the surface-modified microparticle, and any intermediates there between, if any, may be and/or have one or more of the following characteristics: spherical as defined herein, free of covalent crosslinking, free of hydrogel and/or swelling, and have a polydisperse or, preferably, monodisperse, size distribution.
  • the preformed microparticle may be free of lipids and/or encapsulation.
  • the surface-modified microparticle is capable of controlled release, especially sustained release, of the active agent, with a non-limiting release profile such as an initial burst and a linear release profile, and may be provided as a suspension or a dry powder in compositions or formulations for pharmaceutical, therapeutic, diagnostic, cosmetic, and/or nutritional applications.
  • the controlled release may occur within a selected pH environment.
  • the controlled release may occur within a pH range of approximately 2 to 10, and more preferably approximately 5 to 7.5, such as a physiological pH of 7 to 7.4 or endosomal pH of 5 to 6.5.
  • Controlled deposition of the one or more monolayers may further involve alteration of the net surface charge of the microparticle (such as the preformed microparticle onto which one or more of the monolayers have been deposited) through a controlled manipulation of one or more conditions, such as changes in temperature, pressure, pH, ionic strength and/or osmolality of the reaction medium, concentrations of components within the reaction medium, the likes thereof, as well as rates of such changes, and combinations of two or more thereof.
  • Such controlled manipulations may desirably be applied before and up to the deposition of the one or more monolayers, or even during the monolayer formation.
  • the net surface charge of the microparticle is capable of being positive, neutral, and negative.
  • the net surface change is selected through, for example, a controlled change in one or more of the conditions described above, such as a controlled change in pH.
  • the pH of the solution is selected such that the net surface charge of the microparticle is negative, and the difference between the pH of the solution and the surface-neutral point of the microparticle is less than 0.3, alternatively equal to or greater than 0.3, preferably 0.5 or greater, more preferably 0.8 or greater, most preferably 1 or greater.
  • Fig. 1 illustrates one exemplary method of providing surface-modified microparticles in accordance with the present disclosure, is A suspension of a plurality of preformed microparticles used as three-dimensional substrates for the deposition is first provided.
  • Non-limiting methods of forming the preformed microparticle include those disclosed herein and any other methods known to those of skill in the art.
  • One such method is illustrated on the top portion of Fig. 24, which involves providing a solution containing the active agent and the phase-separation enhancing agent, inducing a liquid- solid phase separation through, for example, controlled cooling, and forming the preformed microparticle.
  • any one, two, or more, or all of the compounds used to form the preformed microparticles may preferably be distributed homogeneously throughout each preformed microparticle (e.g., being present at similar concentrations in the center, on the surface, and anywhere else therein).
  • the preformed microparticle may be separated from the liquid phase and, optionally, washed, preferably in the presence of the phase-separation enhancing agent.
  • the washing medium may be the same solution used during phase separation, containing the phase-separation enhancing agent.
  • the preformed microparticle is not separated from the liquid phase or washed.
  • the suspension or a re-suspension of the preformed microparticle is combined and mixed with a solution that includes at least one suitable charged compound.
  • the preformed microparticles may have a weight percent (wt. %) loading of the active agent of 40% or more, preferably 60% or more, or 80% or more, or 90% or more, or 95% or more, and less than 100%, typically 98% or less.
  • the preformed microparticles may further have, or are capable of being induced (such as from a neutral state) to have, a net surface electric charge.
  • the net surface charge is contributed primarily or essentially by the active agent and/or the carrier, if any, present in the preformed microparticles; the compound(s) may preferably be homogeneously distributed therein.
  • the active agent is compartmentalized in one or more portions of the preformed microparticle, such as a center or an underlying layer (a charged monolayer, for example), preferably distributed substantially homogeneously within the portion or primarily on an outer surface thereof.
  • the preformed microparticles may be exposed to (such as mixed with) at least one charged compound having or capable of having a net electrical charge that is, preferably, opposite in sign to the net surface charge of the preformed microparticle, thereby forming the formed monolayer of the charged compound about the preformed microparticle.
  • the formed monolayer or the surface modified microparticle has a net surface electric charge that may be the same in sign as that of the preformed microparticle, zero or, preferably, opposite in sign to that of the preformed microparticle.
  • the formed monolayer may preferably have on its outer surface a positive net surface charge.
  • the formed monolayer may preferably have a negative net surface charge.
  • Deposition of the monolayer can take place in an aqueous medium (e.g., water, buffer, or aqueous solution containing some water miscible organic solvent of the type previously described, or one that may be present in the manufacture of the preformed microparticle).
  • a non-limiting method includes pre-forming or otherwise providing the unmodified microparticle, exposing it to the one or more charged compounds, which may be provided in a solution into which the microparticle -may be immersed, and forming the monolayer.
  • the solution may contain ⁇ one or more of water, a buffer, and a water-miscible organic solvent, and one or more solubility reducing agents (e.g., alcohols, carbohydrates, non-ionic aqueous-miscible polymers, and/or inorganic ionic compounds containing monovalent or polyvalent cations), with a concentration in weight-to-volume percentage of 5% to 50%, preferably 10% to 30%.
  • solubility reducing agents e.g., alcohols, carbohydrates, non-ionic aqueous-miscible polymers, and/or inorganic ionic compounds containing monovalent or polyvalent cations
  • a non-limiting example of the solution contains about 16% (w/v) polyethylene glycol and 0.7% (w/v) NaCl.
  • the pH of the solution typically in a range of 4 to 10, may be adjusted to be same as or close to the surface-neutral point of the core microparticle (such as with a difference of 0 to less than 0.3), or away from that (such as with a difference of 0.3 pH units or greater).
  • the charged compound may be present in the solution at a concentration of 0.05 mg/mL to 10 mg/mL.
  • the preformed microparticle and the charged compound are co-incubated in the solution, preferably at a temperature of 2°C to 5°C or up to ambient temperature over a period of 1 second to 10 hours.
  • the formation of the monolayer may be carried out in a controlled manner.
  • the resulting surface- modified microparticle or an intermediate thereof may be separated from the solution with optional washing.
  • the washing medium may be the same as the solution described above.
  • the procedure may be repeated using alternatingly charged compounds to form the alternatingly charged monolayers, if desired.
  • the reaction system can include one or more solubility reducing and/or viscosity increasing agents (SRA/VIA), as well as one or more PSEAs.
  • Suitable SRA/VIAs and PSEAs include, without limitation, those known to one skilled in the art and those disclosed herein, such as alcohols (e.g., ethanol, glycerol), carbohydrates (such as sucrose), non-ionic aqueous-miscible polymers (e.g., PEG, PVP, block copolymers of polyoxyethylene and polyoxypropylene (poloxamers), hetastarch, dextran, etc.), and inorganic ionizable compounds containing polyvalent (e.g., divalent, trivalent) cations (e.g., metal and organic cations such as those disclosed herein), such as ZnCl 2 .
  • alcohols e.g., ethanol, glycerol
  • carbohydrates such as sucrose
  • non-ionic aqueous-miscible polymers e.g., PEG, PVP, block copolymers of polyoxyethylene and polyoxypropylene (poloxamers),
  • deposition of the formed monolayer may take place in a solution that includes buffered saline (that is, 0.7% NaCl buffer) and 8% or more by weight or volume of a SRA/VIA such as PEG, preferably 12% or more, more preferably 15% or more; typically 30% or less, preferably 25% or less, more preferably 20% or less, most preferably about 16% or more.
  • a SRA/VIA such as PEG
  • the amount of SRA/VIA required in the solution will depend, in part, on the stability of the active agent, as well as the dissolution profile of the monolayer(s).
  • Certain charged compounds (such as the poly cations gelatin B and chitosan) may work in solutions containing 16% or less SRA/VIA.
  • the pH of the solution at which the net surface charge of the microparticle is zero is referred herein as the surface-neutral point of the microparticle in the particular solution.
  • the pH of the solution may be adjusted to be at or near the surface-neutral point of the microparticle in the solution, with a difference there between of less than 0.3 (pH units), preferably 0.25 or less, more preferably 0.2 or less.
  • the pH of the solution may preferably be adjusted away from the surface-neutral point of the microparticle in the solution, with a difference there between of 0.3 (pH units) or . greater, preferably 0.5 or greater, more preferably 0.8 or greater, most preferably 1 or greater.
  • Incubation of the microparticles in the solution can be performed at or, preferably, below ambient temperature, but preferably above the freezing temperature of the solution, to minimize disintegration of the microparticles.
  • Incubation temperature may even be lower than the freezing temperature of the solution when one or more FPDAs disclosed herein are used.
  • the incubation temperature may be between O 0 C and 15 0 C, preferably between 1°C and 10 0 C, more preferably between 2 0 C and 5 0 C, most preferably less than 5°C.
  • the concentration of the charged compound in the solution for each monolayer fabrication may be equal to, less than, and/or greater than one of the following, or in a range between any two thereof: 0.05 mg/mL, 0.1 mg/mL, 0.5 mg/mL, 1 mg/mL, 10 mg/mL, 5 mg/mL, 3 mg/mL.
  • a weight ratio of the preformed microparticle to the charged compound may be 1:1 or greater, preferably 2:1 to 10:1, more preferably 2.5:1 to 7:1
  • Incubation time may be adjusted to achieve the desired charge modification (such as neutralization or charge reversal), monolayer coverage, and/or monolayer thickness. Depending on the particular reaction (such as ingredients and/or conditions), the incubation time may be equal to, shorter than, and/or longer than one of the following, or in a range between any two thereof: 10 hours, 5 hours, 3 hours, 10 minutes, 30 minutes, 100 minutes, 75 minutes, 60 minutes, 15 minutes, 5 minutes, 1 minute, 30 seconds, 10 seconds, 5 seconds, 1 second.
  • Each monolayer may have a thickness that is equal to, less than, and/or greater than one of the following, or in a range between any two thereof: 100 run, 50 nm, 20 nm, 5 nm, 1 nm, 0.5 nm, 0.1 nm, 2 nm, 10 run.
  • a typical monolayer of the present disclosure is less than 100 nm in thickness, preferably less than 10 nm.
  • a factor in controlling release of the active agent from the microparticles may be the type and/or degree of interaction and/or association(e.g., non-covalent association, ionic complexation) that occurs at or near the outer surface of the preformed microparticle (such as the interface with the formed monolayer), which may involve the active agent, the charged compound, and/or other components, if any.
  • a strong interaction or association at this interface slows down, delays, and/or otherwise hinders dissolution of the active agent, and is believed to stabilize the surface-modified microparticle and facilitate fabrication of additional alternatingly charged monolayers, if desired.
  • the interaction can be further affected by the subsequent formation of additional alternatingly charged monolayers.
  • co-incubation of the preformed microparticles 10 and a charged compound 20, preferably in a solution results in intermediate microparticles 40 with a single monolayer of the charged compound 20 formed on and associated with at least the outer surface of the preformed microparticle 10.
  • the suspension of the intermediate microparticles 40 may then be separated from the solution through centrifugation, filtration, diafiltration, and/or other separation methods.
  • the intermediate microparticles 40 are optionally washed with a washing solution (preferably an aqueous medium, such as the SRA-containing buffer described above, as generally shown in Fig. 1).
  • a washing solution preferably an aqueous medium, such as the SRA-containing buffer described above, as generally shown in Fig. 1.
  • the temperature during the incubation and the optional washing is optimized based on the solubility of the active agent and the charged compound 20.
  • the intermediate microparticle 40 may be further exposed to (such as mixed with) a different charged compound 30, preferably in a solution, to form a subsequent monolayer of the charged compound 30 about and associated with the formed monolayer of the intermediate microparticle 40.
  • Charged compound 30 preferably has a net electric charge opposite in sign to that of the charged compound 20.
  • the subsequent monolayer may be formed immediately about the formed monolayer.
  • the intermediate microparticle 50 may have a net surface electric charge that is same in sign as that of the intermediate microparticle 40, neutral or, preferably, opposite in sign to that of the intermediate microparticle 40. As shown in Fig.
  • the monolayer formation procedure may be repeated as in the previous cycle, to form microparticles 50 and 60 that have additional and, preferably but not necessarily, adjacent alternatingly charged monolayers, each associated with the preceding monolayer (Fig. 2).
  • the total number of the monolayers to be formed may be selected or predetermined such that controlled release of the active agent with a desired release profile is achievable in the surface-modified microparticle. As set forth above, this number can be an integer of 1, 2, 3, 4, 5, 6, 7, or greater, preferably 100 or less, more preferably 20 or less, and most preferably 10 or less.
  • one or more of the charged compounds forming the monolayers may be active agent(s) identical to or different from the one in the preformed microparticle.
  • one or more of the odd numbered (e.g., first, third) monolayers may independently be formed of the same or different charged active agent(s), having net electric charge(s) opposite in sign to the net surface charge of the preformed microparticle.
  • one or more of the even numbered (e.g., second, forth) monolayers may independently be formed of the same or different charged active agent(s), having net electric charge(s) same in sign as the net surface charge of the preformed microparticle.
  • charged compound 20 or 30 may be an active agent that is the same as or different from the one in the preformed microparticle 10
  • charged compound 30 or 20, respectively may be an otherwise inert charged compound or a charged active agent different from the one in the preformed microparticle.
  • one or more active agents, charged and/or uncharged may be incorporated into one or more of the monolayers through covalent and/or non- covalent associations.
  • Such monolayer-bound active agent(s) may be the same as that of the preformed microparticle, or different therefrom.
  • Such a construction may allow controlled release (e.g., extended release, sustained release) of the monolayer-bound active agent(s).
  • one of more of such monolayer-bound active agent(s) may be affinity molecules, such as targeting ligands, which may selectively bring the underlying microparticle to a predetermined region to achieve targeted delivery of the active agent within the core microparticle.
  • affinity molecules such as targeting ligands
  • the surface-modified microparticles described above having one or more monolayers of charged compounds, preferably in a suspension may undergo one or more physical and/or chemical treatments to further modify one or more characteristics of the surface-modified microparticles, such as, but not limited to, the release profile of the active agent therein.
  • the treatments may be carried out immediately after the formation of the surface-modified microparticles and prior to any optional washing, or immediately following the optional washings.
  • the treatment may involve manipulation of one or more parameters of the reaction mixture, such as, without limitation, temperature, pH, and/or pressure.
  • the one or more parameters may be adjusted (such as increased or decreased) from an initial value to a second value and held for a period of time, and then adjusted (such as decreased or increased) to a third value or returned or allowed to return to the initial value and held for another period of time.
  • the thermal treatment may involve a heating stage and a cooling stage. Prior to the additional treatment, the suspension may be kept at a relatively low temperature below ambient temperature to at least minimize dissolution of the microparticles therein, preferably the temperature at which the surface-modified microparticles are formed, more preferably 2°C to 10 0 C, such as 4°C.
  • the suspension may be heated to a temperature and incubated at this elevated temperature for an incubation period of 1 minute to 5 hours, preferably 15 minutes to 1 hour, such as 30 minutes.
  • the elevated temperature may be higher than the relatively low temperature at which the suspension was kept prior to the additional treatment, and lower than a degradation temperature of the surface-modified microparticles in the suspension, preferably between 5°C and 40°C, more preferably between 10°C and 30°C.
  • the heating stage may optionally be immediately followed with a cooling stage, during which the suspension may be chilled at a temperature, rapidly or gradually in a controlled manner and optionally incubated at this depressed temperature for an incubation period of 1 minute to 5 hours, preferably 15 minutes to 1 hour, such as 30 minutes.
  • chilling is achieved by washing with a chilled washing solution.
  • the suspension may be allowed to return to or close to its original temperature or to a selected temperature below the temperature to which the suspension was heated.
  • the depressed temperature may be lower than the elevated temperature, and higher than a freezing temperature of the suspension, preferably at or below ambient temperature ⁇ optionally equal to or different from the relatively low temperature at which the suspension was kept prior to the additional treatment, more preferably 15°C or lower, most preferably 10°C or lower, such as 4°C.
  • the resulting mixture may further undergo optional washings as described herein to yield additionally treated, surface-modified, microparticles.
  • Surface-modified microparticles suitable for the additional treatment described above include those formed from amorphous, solid, and homogenous preformed microparticles having 40% to less than 100%, or more typically 80% or greater, by weight, of an active agent as described herein.
  • suitable suspensions include microparticles (such as insulin microspheres) in a buffer, such as a PEG buffer containing 16% PEG, 0.7% NaCl, 67mM Na acetate, and having a pH in the range of 5 to 8 (e.g., 5.7, 5.9, 6.5, 7.0).
  • the microparticles may have a concentration in the buffer of 0.01 mg/ml to 50 mg/ml, preferably 0.1 mg/ml to 10 mg/ml, such as 1 mg/ml.
  • a charged compound or a mixture of two or more thereof, such as protamine sulfate, poly-L-lysine, and/or poly-L-arginine, may be mixed into the suspension to provide a concentration of 0.01 mg/ml to 10 mg/ml, preferably 0.1 mg/ml to 1 mg/ml, such as 0.3 mg/ml.
  • the mixture may be incubated at the relatively low temperature, such as 4 0 C, and under agitation for an incubation period of 10 seconds to 5 hours, such as 1 hour, to ensure the formation of a monolayer of the charged compound on the outer surface of each of the preformed microparticles. Then the suspension may be subjected to the thermal treatment as described above. Optional washings may be carried out on the suspension prior to the additional treatment.
  • the additional treatments may be carried out immediately after the formation of any one or more of the monolayers as disclosed herein.
  • the additional treatment may be carried out immediately after the formation of a single monolayer on the preformed microparticles, the monolayer being formed of positively charged compounds or negatively charged compounds.
  • the additional treatment may or may not be carried out immediately following the formation of such additional monolayers.
  • two or more monolayers may be formed sequentially on the core microparticles, and the additional treatment may be carried out only immediately after a single predetermined monolayer (such as the last monolayer, the first monolayer, or any other monolayer there between) is formed.
  • two or more monolayers may be formed sequentially on the core microparticles, and the additional treatment may be carried out immediately after the formation of each and every monolayer having one or more predetermined characteristics, such as containing positively charged or negatively charged compounds, or containing a particular compound (e.g., active agent, affinity molecule, derivative )or moiety (e.g., functional group, label), or being a particular monolayer from the core (e.g., first, second, third, fourth, fifth), hi a further example, the additional treatment may be carried out immediately after the formation of each monolayer of a predetermined set, which may be all of the monolayers or a subset thereof.
  • a predetermined set which may be all of the monolayers or a subset thereof.
  • the surface-modified microparticles following the additional treatment may display modifications in net surface charge (zeta potential) and/or release profile of the active agent therein.
  • zeta potential net surface charge
  • release profile of the active agent therein.
  • certain charged compounds such as PLL and PLA 5 but not protamine sulfate
  • a change such as an increase
  • the additionally treated, surface-modified microparticles are capable of displaying a reduction in the 1- hour percentage of cumulative release (%CRi h ) of the active agent therein, as compared to the surface-modified microparticles without the additional treatment.
  • the example demonstrates that the initial burst of the active agent release may be significantly reduced by the additional treatment.
  • surface- modified microparticles may be capable of continued, preferably sustained, release beyond 1 hour, preferably beyond 24 hours, more preferably beyond 48 hours, most preferably beyond 7 days, having a 24-hour percentage of cumulative release (%CR 24h ) that is greater than %CRi h .
  • the surface-modified microparticles of the present disclosure when subjected to in vitro release in a release buffer (10 mM Tris, 0.05% Brij 35, 0.9% NaCl, pH 7.4, free of divalent cation) at 37°C, may be capable of displaying a %CRu, of 50% or less and/or a ratio of %CR 24h to %CR 1 h of greater than 1:1.
  • the %CRi h may preferably be 40% or less, more preferably 30% or less, further preferably 20% or less, most preferably 10% or less.
  • the ratio of %CR 24h to %CR 1h may preferably be 1.05:1 or greater, more preferably 1.1:1 or greater, but not more than 10:1, preferably 5:1 or less, more preferably 2:1 or less, most preferably 1.5:1 or less.
  • the additional treatment following the monolayer formation as disclosed herein allows the charged compound in the monolayer and the molecules (e.g., the active agent, the optional carrier molecules in the preformed microparticle, the charged compound in the preceding monolayer) that comprises the outer surface of the substrate (e.g., the preformed microparticle, the preceding monolayer) to rearrange and form an association that is much stronger than the electrostatic interaction between the monolayer and the outer surface of the substrate prior to the additional treatment. It is believed that through the additional treatment a modified shell is formed on the outer surface of the surface-modified microparticle, the modified shell containing a homogenous mixture of the charged compound and the molecules that form the outer surface of the substrate.
  • the molecules e.g., the active agent, the optional carrier molecules in the preformed microparticle, the charged compound in the preceding monolayer
  • the substrate e.g., the preformed microparticle, the preceding monolayer
  • Deposition of additional alternatingly charged monolayers of charged compounds beyond the formed monolayer may further affect, among other things, the release profile of the active agent in the preformed microparticle. As previously described, depending on the attractive forces at the interface between the preformed microparticle and the formed monolayer, strong association between the two may be observed. This may result in retarding the quantity and/or rate of release of the active agent.
  • the release profile may be further modified by forming one or more additional alternatingly charged monolayers about the formed monolayer. Without being restricted to any particular theory, it is believed that addition of a second oppositely charged monolayer may ease the association between the formed monolayer and the preformed microparticle, thereby enhancing the release of the active agent.
  • Suitable charged compounds that may be used in accordance with the present invention may be charged compounds capable of associating with any substrate, preferably by, but not limited to non-covalent association and, more preferably, electrostatic interaction.
  • suitable charged compounds include positively charged, negatively charged, or zwitterionic, and include, but are not limited to, polyelectrolytes, charged polyaminoacids, polysaccharides, polyionic polymers, ionomers, charged peptides, charged proteinaceous compounds, charged lipids optionally in combination with uncharged lipids, charged lipid structures such as liposomes, precursors and derivatives thereof, and combinations of two or more thereof.
  • Non-limiting examples include negatively charged polyelectrolytes such as polystyrene sulfonate (PSS) and polyacrylic acid (PAA), negatively charged polyaminoacids such as polyaspartic acid, polyglutamic acid, and alginic acid, negatively charged polysaccharides such as chondroitin sulfate and dextran sulfate, positively charged polyelectrolytes such as polyallyl amine hydrochloride (PAH) and poly(diallyldimethyl ammonium chloride (PDDA), positively charged polyaminoacids such as poly(L-lysine) hydrochloride, polyornithine hydrochloride, and polyarginine hydrochloride, and positively charged polysaccharides such as chitosan and chitosan sulfate.
  • PES polystyrene sulfonate
  • PAA polyacrylic acid
  • negatively charged polyaminoacids such as polyaspartic
  • biocompatible polyionic polymers e.g., ionomers, polycationic polymers such as polycationic polyurethanes, polyethers, polyesters, polyamides; polyanionic polymers such as polyanionic polyurethanes, polyethers, polyesters, polyamides), charged proteins (e.g., protamine, protamine sulfate, xanthan gum, human serum albumin, zein, ubiquitins, and gelatins A & B), and charged lipids (e.g., phosphatidyl choline, phosphatidyl serine).
  • biocompatible polyionic polymers e.g., ionomers, polycationic polymers such as polycationic polyurethanes, polyethers, polyesters, polyamides
  • polyanionic polymers such as polyanionic polyurethanes, polyethers, polyesters, polyamides
  • charged proteins e.g., protamine, protamine sulfate, x
  • derivatives e.g., glycosylated, hyperglycosylated, PEGylated, FITC- labeled, salts thereof
  • conjugates and complexes of the charged compound disclosed herein.
  • suitable positively charged lipids that is, polyanionic lipids
  • negatively charged lipids that is, polycationic lipids
  • zwitterionic lipids include 1,2- distearoyl-sn-glycero-3 -phosphocholine (DSPC), 1 ,2-dioleoyl-sn-glycero-3 - - - -
  • lipid structures such as liposomes
  • Uncharged (such as non-ionic) lipids may be used in combination with electrically charged lipids to form one or more of the monolayers, and the molar ratio there between can be optimized to achieve minimum permeability of the active agent through the monolayer.
  • the surface-modified microparticle disclosed herein typically containing a preformed microparticle and one or more monolayers, preferably has a release profile of the active agent that is different from that of the core microparticle.
  • the differences in release profile include a reduction in the initial burst, an extension of release time, a display of linear/constant release over a time period, and/or a reduction in rate of release over a prolonged time period.
  • the surface-modified microparticles may be present, preferably in a functionally (e.g., therapeutically, pharmaceutically, diagnostically) effective amount, as a suspension or dry powder in a liquid or solid composition or formulation, in the presence or absence of one or more of preservatives, isotonicity agents, pharmaceutically acceptable carriers, and stabilizing agents.
  • Such compositions and formulations may be administered in an effective amount to a subject for prevention or treatment of a condition or state, or as a nutritional supplement, or for the purpose of physical enhancement or psychological well-being
  • Such compositions and formulations may be incorporated into a diagnostic method, tool, or kit for in vitro and/or in vivo detection of a substance, condition, or disorder being present or absent, or a disposition for such a condition or disorder.
  • the substance upon contact, may form an association (e.g., conjugate, complex) with the surface-modified microparticle or a portion thereof (such as the core microparticle), which is capable of providing one or more signals for detection.
  • the one or more signals may be one or more moieties labeled on one or more portions of the association (e.g., the substance, the microparticles), or may be elicited upon the formation of the association (e.g., emission of light, discharge of another substance).
  • the surface-modified microparticles may be incorporated into a nutritional and/or dietary supplement or a food composition, or used as a food additive, for prevention and/or treatment of a condition or disorder in a subject.
  • the QCM method was used to confirm the presence of layer-by-layer assembly of electrically charged compounds in the presence of an SRA-containing solution.
  • Precursor films of multiple (e.g., 2, 3, or 4) PAH/PSS bilayers i.e., each bilayer includes a PAH monolayer immediately adjacent to a PSS monolayer
  • QCM USI QCM System, Model 260303, Sanwa Tsusho Co., Ltd, Japan
  • each monolayer the resonators were incubated in 0.25 M NaCl aqueous buffer with a charged compound concentration of 1.5 mg/mL at room temperature for 15 minutes, washed three times with deionized (DI) water, and dried.
  • DI deionized
  • the monolayers can also be formed using DI water solution having a charged compound concentration of 3 mg/mL.
  • the coated resonators were further incubated in aqueous buffers containing the respective charged compounds at +2°C for about 1 hour, followed by washings with DI water.
  • concentration of the charged compound in the buffer was in a range of 0.1 mg/mL to 3 mg/mL, preferably 1 mg/mL.
  • concentration of the charged compound in the buffer was 0.1 mg/mL to 3 mg/mL, preferably 0.25 mg/mL to 1 mg/mL.
  • zeta potential microparticle net surface charge
  • a Zeta Potential Analyzer Model ZetaPALS, Brookhaven Instruments Corp., Holtsville, NY
  • a 40 ⁇ L aliquot of each sample under investigation was added to 1.5 niL of the corresponding salt-free PEG solution, mixed, and the resulting suspension was immediately used for the measurements.
  • the temperature of the suspension was equilibrated to 8°C to minimize microparticle disintegration.
  • a 10 mL aliquot of a releasing buffer (10 mM Tris, 0.05% Brij 35, 0.9% NaCl, pH 7.4) was added into a glass vial containing 0.5 mL of the concentrated particle suspension (equivalent to 3 mg of insulin), mixed, and incubated at 37°C.
  • a releasing buffer 10 mM Tris, 0.05% Brij 35, 0.9% NaCl, pH 7.4
  • the concentrated particle suspension equivalent to 3 mg of insulin
  • 400 ⁇ L of the rVR medium was transferred into a microfuge tube and centrifuged for 2 minutes at 13k rpm.
  • a 300 ⁇ L aliquot of the supernatant was removed and stored at -80°C until analyzed by Bicmchoninic Acid (BCA) assay as understood by one skilled in the art.
  • BCA Bicmchoninic Acid
  • Example IA Microspheres with Polystyrene Sulfonate Monolayer
  • Insulin microspheres formed using a phase separation method disclosed herein i.e., preformed microparticles
  • a phase separation method disclosed herein i.e., preformed microparticles
  • aqueous solution of 16% (w/v) PEG and 0.7% (w/v) NaCl in the presence of 0.3 mg/mL polyion PSS at 2 0 C and pH 4.8 for 1 hr.
  • To remove the unassociated PSS centrifugal washing (at 3000 rpm for 15 minutes) was applied twice, each using the initial volume of the aqueous solution described above as the washing medium to re-suspend the microspheres.
  • Comparison of zeta-potential values of the unmodified and modified microspheres confirms the formation of the PSS monolayer (FIG. 3).
  • Example IB Microspheres with Multiple Alternating Monolayers of Polystyrene Sulfonate and Polyallylamine Hydrochloride
  • Example IA The PSS-modified microparticles of Example IA were used as intermediate microparticles to form a subsequent monolayer of polycation PAH. Similar formation and washing procedures were used as described in Example IA 5 except that PAH was substituted for PSS at the same concentration. The procedures were repeated to form desired number of alternating monolayers.
  • FIG. 3 illustrates the zeta-potential values of four consecutive depositions of the PSS/PAH bilayer assembly after formation of each monolayer.
  • Example 2A Microspheres Surface-modified at a pH Below the Surface-neutral Point of the Microspheres
  • Example IA The procedures of Example IA were used to form a polyanion monolayer on insulin microspheres at pH 4.8 (below surface-neutral point of the microspheres, which was observed to be about 5.6).
  • FIG. 4 depicts zeta-potential values of insulin microspheres with a monolayer of polyacrylic acid (model polyanion), dextran sufate, polyaspartic acid, polyglutamic acid, and alginate.
  • the zeta-potential of the preformed insulin microspheres at pH of 4.8 showed a positive net surface charge.
  • the net surface charges of the resulting surface-modified microspheres were negative.
  • Example 2B Microspheres Surface-modified at a pH Above the Surface-neutral Point of the Microspheres.
  • FIG. 5 depicts zeta-potential values of insulin microspheres with a monolayer of polydiallyldimethylammonium chloride (PDDA, model polycation), protamine sulfate (ProtS), poly-1-arginine (PLA), and poly-1-lysine (PLL).
  • PDDA polydiallyldimethylammonium chloride
  • ProtS protamine sulfate
  • PLA poly-1-arginine
  • PLL poly-1-lysine
  • microspheres of Examples IA & 2A were used as intermediate microspheres, re-suspended in the aqueous solution (16% PEG, 0.7% NaCl, pH 4.8) in the presence of 0.3 mg/mL polycation PLL, and incubated for 1 hr at 2 0 C to form a subsequent monolayer of PLL over the formed polyanion monolayer.
  • a QCM as described above was used to measure the thickness of each polyion monolayer.
  • the reaction medium contained PLL or chondroitin sulfate at 1 mg/mL in 16% PEG, 0.7% NaCl.
  • the film assembly was constructed by consecutive incubation of the QCM resonators in the reaction media containing the polyions for 15 minutes each, followed intermediately with DI water washes and drying with nitrogen stream.
  • FIG. 9 illustrates the progressive film thickness following formation of each monolayer. Depending on the polyion, each monolayer was estimated to increase the total thickness by about 1 run or less, with an averaged increase of about 0.5 nm.
  • Example 3B Microspheres with Multiple Monolayers of Oppositely Charged Biocompatible Polyions
  • Condroitin sulfate and gelatin A were used to form multiple monolayers of oppositely charged polyions about preformed insulin microspheres, in an aqueous solution of 16% PEG & 0.7% NaCl, at pH 4.8 and 2 0 C. Formation of the condroitin sulfate monolayer, according to the procedures of Example IA, was followed by subsequent formation of the gelatin A monolayer. The procedures were repeated to form a total of 6 alternatively charged monolayers. Reversal of net surface charge of the microspheres following formation of each monolayer is shown in FIG. 10.
  • Example IB The procedures of Example IB were used to form monolayers of protamine and condroitin sulfate about preformed insulin microspheres, with the exception that the aqueous solution had a pH of 6.4 and contained 16% PEG, 0.7% NaCl, 0.16% (w/v) acetic acid, and 0.026% (w/v) ZnCl 2 . Comparative examples were formed using a Zn-free & acetate-free aqueous solution with a pH of 6.4 containing 16% PEG and 0.7% NaCl. Zeta- potentials of the resulting microspheres are shown in FIG. 11.
  • Example IB The procedures of Example IB were used to form monolayers of protamine and condroitin sulfate about preformed insulin microspheres, with the exception that the aqueous solution was PEG-free, had a pH of 7.0, and contained 0.7% NaCl, 0.16% acetic acid, and 0.026% ZnCl 2 . Zeta-potentials of the resulting microspheres are shown in FIG.
  • DAP cationic lipid l,2-dioleoyl-3- dimethylammonium-propane
  • DOPC zwitterionic l,2-dioleoyl-sn-glycero-3- phosphocholine
  • Example 4B 0.16% acetic acid and 0.026% ZnCl 2 at pH 7.0 were used to co-incubate with preformed insulin microspheres (pre-washed with the same aqueous solution) at 2°C for 1 hr.
  • the procedures of Example 4B were applied to form a subsequent monolayer of chondroitin sulfate.
  • FIG. 13 illustrates LSC micrographs of microspheres containing Rodamine B-labeled protamine monolayer (top right), microspheres containing FITC-labeled DAP (top left), and microspheres containing both (bottom left). Zeta-potential values of the microspheres following each deposition are shown in FIG. 14.
  • Protamine-modified insulin microspheres were formed using the procedures of Example 2B at various concentrations of the polyion in the reaction medium.
  • the IVR profiles of resulting protamine-modif ⁇ ed insulin microspheres are shown in FIG. 15. An increase in concentration of polyion reduced the initial burst and subsequent release rate of insulin from the surface-modified microspheres.
  • Example 7 Release Modification with Multiple Monolayers
  • the resulting microspheres of Example 2B were used as intermediate microparticles, about which carboxymethyl cellulose (CMC) was deposited at various concentrations in the reaction medium.
  • CMC carboxymethyl cellulose
  • the IVR profiles shown in FIG. 16 demonstrated the capability of the subsequent monolayer in further modifying the release of active agents from the surface-modified microspheres. Deposition of an additional protamine monolayer was able to partially or fully restore the release profile (FIG. 17).
  • Example 8 In Vivo Release of Insulin from Protamine-modified Insulin Microspheres [000168] In vivo release of insulin from protamine-modified insulin microspheres was investigated in chemically induced Sprague-Dawley rats.
  • the surface-modified microspheres prepared according to Example 2B were administered as a suspension in 16% PEG 3350, pH 7.0.
  • the preformed insulin microspheres free of surface modification were administered in the PEG solution, or in phosphate-buffered saline, pH 7.4, as control.
  • the animals received an initial subcutaneous dose of 1 IU/kg of the microspheres.
  • An ELISA assay was used to determine the recombinant human insulin (rhlNS) serum levels in the collected samples.
  • the surface-modification increased the maximum serum concentration of rhlNS (C max ) and the time to achieve C max (t max ), as well as the area under the rhlNS concentration-time curve (AUC) and the mean residence time (MRT) of the protein.
  • the serum glucose depression (FIG. 18B) also was in agreement with the corresponding serum rhlNS.
  • the C max of the surface modified microparticle was 2.5 fold greater than the C max of the preformed microparticle.
  • the C max of the surface modified microparticle may be increased 1.1 fold or greater, 1.25 fold or greater, 1.5 fold or greater, 2.0 fold or greater than the C max of the preformed microparticle.
  • Aqueous media of protamine sulfate (0.15 mg/mL) used to incubate the preformed insulin microparticles contained one of PLURONIC® F-68 or F-127 (10% or 16% w/v), glycerol (20%, 40%, or 60% v/v), and ethanol (10% v/v). Procedures as described in Example IA were followed. Zeta-potential values of the microspheres before and after surface modification, shown in FIG. 19, indicated formation of protamine monolayer.
  • Example IA Procedures of Example IA were followed, in which the concentration of protamine sulfate was varied in a range of 0.1 mg/mL to 1.5 mg/mL.
  • FIG 2OA illustrates the relationship between zeta-potential of the microspheres and the cumulative release of insulin at 48 hrs. An increase in protamine concentration in the reaction mediumcorrelated with reduction of insulin release, with an observed maximum effective concentration of about 0.3 mg/mL.
  • protamine-modified microspheres prepared at a protamine concentration of 1.5 mg/mL were further modified with polyanion carboxymethyl cellulose or chondroitin sulfate in the concentration range of 0.05-1.2 mg/mL or 0.1-1.2 mg/mL, respectively.
  • Formation of the subsequent monolayer significantly reversed the release reduction effect of the protamine monolayer, as shown in FIGs. 2OB and 2OC.
  • Example 11 Surface Modification of hGH Microspheres
  • Preformed hGH microspheres were incubated, in alternating sequence, in aqueous media (16% PEG 3350, 0.7% NaCl, pH 6.0) containing 0.3 mg/mL protamine sulfate and chondroitin sulfate, respectively, at 2 0 C for 1 hr each, to form the alternatingly charged monolayers.
  • FIG 21 A depicts the zeta-potential of the microspheres after deposition of each monolayer.
  • Example 12 Surface Modification of Microspheres of Intravenous Immunoglobulin
  • IVIG intravenous immunoglobulin
  • Preformed intravenous immunoglobulin (IVIG) microspheres were modified with alternating monolayers of chondroitin sulfate and protamine sulfate.
  • the incubation was carried out in a pH 7.0 aqueous medium containing 12.5% PEG 8000, 50 mM ammonium acetate, and 0.15 mg/mL of the respective polyion at 4°C for 1 hr. Centrifugal washing was used to remove excess polyions.
  • FIG 22 depicts the zeta-potential of the microspheres after deposition of each monolayer.
  • Example 13 Surface Charge Characteristics of Microspheres in Aqueous PEG Media
  • pH of the media was adjusted in a range of 4-7.5. Zeta-potential of the microspheres were determined in each medium, and plotted versus the corresponding pH, as shown in FIG. 23.
  • the surface- neutral point for the preformed insulin microspheres was estimated to be 5.6. As the pH of the medium decreased below or increased above the surface-neutral point, the net surface charge of the preformed insulin microspheres became more and more positive or negative, respectively.
  • insulin microspheres (20 mg) formed using a phase separation method disclosed herein were suspended in 19 ml of a buffer [containing 16% (w/v) PEG, 0.7% (w/v) NaCl, and 67 mM sodium acetate] at one of the following pH values: 5.7, 5.9, 6.5, and 7.0.
  • Zeta potential of the unmodified microspheres in the buffer of different pH was measured as described herein above.
  • Protamine sulfate, poly-1-lysine, or poly-1- arginine was added to the suspension as 1 ml of a 6 mg/ml solution in the same buffer at the same pH as that of the suspension.
  • the resulting reaction mixtures each had a microsphere concentration of 1 mg/ml and a polycation concentration of 0.3 mg/ml.
  • the reaction mixtures were incubated at 4°C for one hour, and then centrifugally washed (3000 rpm for 15 minutes) three times with 20 ml fresh aliquots of the buffer at the respective pH values of the reaction mixtures.
  • Zeta potentials of the resulting surface-modified microspheres in the resuspensions of the last washing were measured as described above.
  • the surface-modified microspheres were then subjected to in vitro release following the protocol disclosed herein.
  • the zeta potential of the PLL-modified insulin microspheres was higher (in a general range of 15 mV or greater, such as about 20 mV) at reaction pH values (e.g., 5.7, 5.9) close to the surface-neural point of the unmodified insulin microspheres (insulin SNP COre5 about 5.6), and lower (in a general range of less than 15 mV, such as about 8 mV) at reaction pH values (e.g., 6.5, 7.0) away from insulin SNP core -
  • the magnitude (about 30 mV) of the charge reversal following the formation of the PLL monolayer was the same across the different reaction pH values specified above.
  • the zeta potential of the PLA-modified insulin microspheres was higher (above 20 mV) at reaction pH values close to insulin SNP core , and lower (below 20 mV) at reaction pH values away from insulin SNP CO r e -
  • the magnitude of the charge reversal following the formation of the PLA monolayer was less (about 30 mV) at reaction pH values close to insulin SNP cor e, and greater (about 40 mV) at reaction pH values away from insulin SNP core -
  • the zeta potential (about 20 mV) of the ProtS-modified insulin microspheres was the same across the different reaction pH values specified above.
  • the magnitude of the charge reversal following the formation of the protamine sulfate monolayer was less (about 30 mV or less) at reaction pH values close to insulin SNP CO re 5 and greater (about 40 mV or greater) at reaction pH values away from insulin SNP core - [000178]
  • the in vitro 1-hour percentage of cumulative release (%CRi h ) of insulin from the surface-modified insulin microspheres was affected by the reaction pH and/or the polycation used in the surface modification reaction. Specifically, insulin %CR 1 h was generally greater at reaction pH values close to insulin SNP cor e than that at reaction pH values away from insulin SNP core , with the difference there between ranging from greater than 5% to 10% to 20% to less than 30%.
  • PLA-modified microspheres and ProtS-modified microspheres had comparable insulin %CR 1h , the level of which was less than that of PLL-modified microspheres, with the differences there between ranging generally from 20% to 30% or more.
  • the preformed microparticles of the present disclosure may be surface-modified at reaction pH away from SNP ⁇ r e using certain charged compounds (e.g., protamine sulfate, PLA).
  • the preformed microparticles of the present disclosure may be surface-modified at reaction pH close to SNP cor e using certain charged compounds (such as PLL).
  • PLL certain charged compounds
  • Nucleic acid microspheres were formed according to the disclosures of U.S. Patent Application Publication Nos. 2006-0018971 and 2006-0024240, the entirety of which are incorporated herein by express reference thereto. Each of the microspheres had a homogenous mixture containing at least 80% (such as 85% to 90%) by weight of a CD40 siRNA and 15% or less (such as 6% to 10%) by weight of poly-1-lysine.
  • the nucleic acid microspheres were suspended in 100 ⁇ l of nuclease-free deionized water (pH 7.0) containing 1 mg/ml Rodamine B-labeled PLL (70 kD).
  • the suspension was incubated with agitation at 4°C for 45 minutes to form the surface-modified microspheres, which were then centrifugally washed with nuclease-free deionized water (pH 7.0).
  • the zeta-potentials of the unmodified microspheres and the surface-modified microspheres were measured to be —24 mV and 34 mV, respectively.
  • surface modification of the nucleic acid microspheres through formation of a polycation monolayer is capable of reversing their surface electric charge.
  • Laser scanning confocal micrograph of Rodamine B-labeled PLL is shown in FIG. 27, confirming successful formation of the monolayer on the outer surface of the nucleic acid microspheres.
  • Example 16 Thermal treatment of surface-modified microparticles
  • a 1.5 ml aqueous solution of a polycation (ProtS, PLL, or PLA) dissolved in the same buffer was mixed with the suspension, resulting in reaction mixtures having a microsphere concentration of 4 mg/ml and a polycation concentration of 3 mg/ml.
  • reaction mixtures were incubated at 4°C with continuous agitation for 30 minutes to form the surface-modified insulin microspheres having the respective polycation monolayer.
  • the reaction mixtures were further incubated for another 30 minutes, at a temperature of 4°C, 15°C, 28°C, or 37 0 C.
  • the thermally- treated insulin microspheres were centrifugally (3000 rpm at 4°C for 10 minutes) collected and washed three times with fresh aliquots of the buffer at pH 7.0 and 4 0 C. Zeta potential data and in vitro release profiles of the thermally-treated, surface-modified insulin microspheres were generated as described herein above.
  • the incubation at different elevated temperatures as described above results in different levels of initial insulin release from the polycation-modified insulin microspheres (insulin %CRlh being lower following incubation at 28°C than that following incubation at 15 0 C), which are consistently less than that following incubation at non-elevated temperature (i.e., 4 0 C).
  • the reduction effect of the thermal treatment (at 28 0 C) on the initial release phase of the in vitro release of insulin was consistently observed in the polycations tested above. Unexpectedly, different polycations exert different levels of reduction on the initial release of insulin. As illustrated in FIGS. 26 and 28, and summarized in Table 3, the reduction effect observed in PLA-modif ⁇ ed insulin microspheres of the present disclosure is greater than that in PLL-modified insulin microspheres, while the reduction effect observed in ProtS-modif ⁇ ed insulin microspheres is less than that in PLL-modified insulin microspheres.
  • Example 17 Thermal treatment of surface-modified microparticles
  • Preformed, unmodified hGH microspheres such as those formed from controlled phase separation as disclosed herein, underwent the same thermal treatments as described in Example 16.
  • Zeta potential data and in vitro release profiles of the thermally- treated, surface-modified hGH microspheres were generated as described herein above.
  • %CRm of the surface-modified hGH microspheres following incubation at 28 0 C was reduced as compared to that of the surface-modified hGH microspheres following incubation at 4°C.
  • Table 4 Percentage of Reduction in %CRu, of PLA-modif ⁇ ed hGH Microspheres
  • Example 18 Effect of Thermally-treated, Surface-modified Microparticles In Vivo [000185]
  • Unmodified insulin microspheres were prepared using the controlled phase separation method disclosed herein. Two portions. of the unmodified insulin microspheres were surface-modified with PLA, with one portion thermally-treated (at 28 0 C), and the other portion incubated at 4 0 C as control, according to the procedures described in Example 16. Injectable compositions each containing one of the three different insulin microspheres suspended in a buffer [16% (w/v) PEG, 0.7% (w/v) NaCl, at pH 7.0] were prepared.
  • compositions were administered subcutaneously to normal Spague-Dawley rats at a dose of 4 IU/kg, An ELISA assay was used to determine the insulin serum levels in the collected samples. The results are illustrated in Table 5 and FIGs. 29A and 29B. As shown in Fig. 29A, CR 1I i was likewise reduced following heat treatment.
  • PLA-modified insulin microspheres treated at 4°C provided comparable serum insulin concentration profile, serum glucose depression profile, C max , and t max .
  • PLA-modified insulin microspheres treated at 28 0 C provided flattened and right-shifted serum insulin concentration profile, right-shifted serum glucose depression profile, depressed C max , and prolonged t max . Table 5.
  • Fig. 29 A is a graph showing the serum insulin concentration versus time profiles in rats that have received a single subcutaneous injection of uncoated insulin microspheres, PLA-modified insulin microspheres treated at 28°C, or PLA-modified insulin microspheres treated at 4 0 C (Example 18);
  • Fig. 29B is a graph showing the serum glucose depression versus time profiles of rats treated with single subcutaneous injection of uncoated insulin microspheres, PLA-modified insulin microspheres treated at 28 0 C, or PLA-modified insulin microspheres treated at 4 0 C (Example 18);

Landscapes

  • Health & Medical Sciences (AREA)
  • Chemical & Material Sciences (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Engineering & Computer Science (AREA)
  • Bioinformatics & Cheminformatics (AREA)
  • Medicinal Chemistry (AREA)
  • General Health & Medical Sciences (AREA)
  • Veterinary Medicine (AREA)
  • Public Health (AREA)
  • Animal Behavior & Ethology (AREA)
  • Pharmacology & Pharmacy (AREA)
  • Epidemiology (AREA)
  • Molecular Biology (AREA)
  • Organic Chemistry (AREA)
  • Biochemistry (AREA)
  • Dermatology (AREA)
  • Biophysics (AREA)
  • Diabetes (AREA)
  • Endocrinology (AREA)
  • Chemical Kinetics & Catalysis (AREA)
  • Genetics & Genomics (AREA)
  • General Chemical & Material Sciences (AREA)
  • Proteomics, Peptides & Aminoacids (AREA)
  • Dispersion Chemistry (AREA)
  • Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
  • Biomedical Technology (AREA)
  • Neurosurgery (AREA)
  • Medicinal Preparation (AREA)
  • Medicines That Contain Protein Lipid Enzymes And Other Medicines (AREA)
  • Glanulating (AREA)
  • Manufacturing Of Micro-Capsules (AREA)

Abstract

L'invention concerne des microparticules à surface modifiée ainsi que des procédés de fabrication et d'utilisation associés. Ces microparticules à surface modifiée comprennent une microparticule préformée ou centrale qui contient au moins un agent actif. La surface extérieure de cette microparticule préformée ou centrale transporte une charge de surface nette. Une monocouche est associée à la surface extérieure de la microparticule préformée ou centrale. Cette monocouche contient au moins un composé chargé présentant une charge différente de la charge de surface nette de la microparticule préformée ou centrale.
EP06751564A 2005-04-27 2006-04-27 Microparticules a surface modifiee et procedes de formation et d'utilisation associes Withdrawn EP1885335A1 (fr)

Applications Claiming Priority (3)

Application Number Priority Date Filing Date Title
US67537205P 2005-04-27 2005-04-27
US75090305P 2005-12-16 2005-12-16
PCT/US2006/015918 WO2006116546A1 (fr) 2005-04-27 2006-04-27 Microparticules a surface modifiee et procedes de formation et d'utilisation associes

Publications (1)

Publication Number Publication Date
EP1885335A1 true EP1885335A1 (fr) 2008-02-13

Family

ID=36928379

Family Applications (1)

Application Number Title Priority Date Filing Date
EP06751564A Withdrawn EP1885335A1 (fr) 2005-04-27 2006-04-27 Microparticules a surface modifiee et procedes de formation et d'utilisation associes

Country Status (8)

Country Link
US (1) US20060260777A1 (fr)
EP (1) EP1885335A1 (fr)
JP (1) JP2008539259A (fr)
CN (1) CN101188996B (fr)
AU (1) AU2006241145B2 (fr)
CA (1) CA2604225A1 (fr)
MX (1) MX2007013356A (fr)
WO (1) WO2006116546A1 (fr)

Families Citing this family (54)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US9006175B2 (en) 1999-06-29 2015-04-14 Mannkind Corporation Potentiation of glucose elimination
ES2425392T3 (es) 2002-03-20 2013-10-15 Mannkind Corporation Cartucho para un aparato de inhalación
DK1786784T3 (da) 2004-08-20 2011-02-14 Mannkind Corp Katalyse af diketopiperazinsyntese
BR122019022692B1 (pt) 2004-08-23 2023-01-10 Mannkind Corporation Composição terapêutica em pó seco contendo dicetopiperazina, pelo menos um tipo de cátion e um agente biologicamente ativo
WO2007033372A2 (fr) 2005-09-14 2007-03-22 Mannkind Corporation Procede de preparation de medicaments fonde sur l'accroissement de l'affinite des surfaces de microparticules cristallines pour des principes actifs
DK1986679T3 (da) 2006-02-22 2017-11-20 Mannkind Corp Fremgangsmåde til forbedring af mikropartiklers farmaceutiske egenskaber omfattende diketopiperazin og et aktivt indholdsstof
DE602007008125D1 (de) 2006-10-31 2010-09-09 Surmodics Pharmaceuticals Inc Kugelförmige polymer-teilchen
US8697098B2 (en) 2011-02-25 2014-04-15 South Dakota State University Polymer conjugated protein micelles
CA3153292A1 (fr) 2008-06-13 2009-12-17 Mannkind Corporation Inhalateur de poudre seche et systeme d'administration de medicament
US8485180B2 (en) 2008-06-13 2013-07-16 Mannkind Corporation Dry powder drug delivery system
CN102065942B (zh) 2008-06-20 2013-12-11 曼金德公司 用于对吸入工作进行实时描绘的交互式设备和方法
US20100009007A1 (en) * 2008-07-10 2010-01-14 Baxter International Inc. Non-covalent modification of microparticles and process of preparing same
TWI532497B (zh) 2008-08-11 2016-05-11 曼凱公司 超快起作用胰島素之用途
US8323685B2 (en) * 2008-08-20 2012-12-04 Baxter International Inc. Methods of processing compositions containing microparticles
US8367427B2 (en) 2008-08-20 2013-02-05 Baxter International Inc. Methods of processing compositions containing microparticles
US8323615B2 (en) 2008-08-20 2012-12-04 Baxter International Inc. Methods of processing multi-phasic dispersions
BRPI0922344A2 (pt) 2008-12-19 2017-10-24 3B Pharmaceuticals Gmbh inibidores de tfpi e métodos de uso
ES2319158B1 (es) * 2008-12-23 2010-01-26 Grifols, S.A Composicion de microparticulas biocompatibles de acido alginico para la liberacion controlada de principios activos por via intravenosa.
WO2010074675A1 (fr) * 2008-12-23 2010-07-01 Board Of Regents Of The University Of Texas System Particules ciblant l'inflammation
US8314106B2 (en) 2008-12-29 2012-11-20 Mannkind Corporation Substituted diketopiperazine analogs for use as drug delivery agents
EP2403490B1 (fr) 2009-03-04 2019-08-07 MannKind Corporation Système amélioré pour la délivrance d'un médicament sous la forme d'une poudre sèche
PL2405963T3 (pl) 2009-03-11 2014-04-30 Mannkind Corp Urządzenie, układ i sposób pomiaru oporu inhalatora
ES2347119B2 (es) 2009-04-22 2011-04-28 Universidad De Santiago De Compostela Nanocapsulas de poliarginina.
KR20180036807A (ko) 2009-06-12 2018-04-09 맨카인드 코포레이션 한정된 비표면적을 갖는 디케토피페라진 마이크로입자
CA2778698A1 (fr) 2009-11-03 2011-05-12 Mannkind Corporation Appareil et methode de simulation d'efforts d'inhalation
EP2496700B1 (fr) 2009-11-04 2017-03-01 The University Of British Columbia Particules lipidiques contenant des acides nucléiques et procédés associés
WO2011115712A2 (fr) 2010-03-19 2011-09-22 Baxter International Inc Inhibiteurs de tfpi et procédés d'utilisation associés
RU2571331C1 (ru) 2010-06-21 2015-12-20 Маннкайнд Корпорейшн Системы и способы доставки сухих порошковых лекарств
CA2828255A1 (fr) 2011-02-25 2012-08-30 South Dakota State University Nanovecteurs de proteines s'appliquant a une administration topique
CA2828253C (fr) 2011-02-25 2016-10-18 South Dakota State University Micelles de proteines conjuguees a un polymere
KR101940832B1 (ko) 2011-04-01 2019-01-21 맨카인드 코포레이션 의약 카트리지용 블리스터 패키지
WO2012174472A1 (fr) 2011-06-17 2012-12-20 Mannkind Corporation Microparticules de dicétopipérazine de capacité élevée
EP2776053A1 (fr) 2011-10-24 2014-09-17 MannKind Corporation Procédés et compositions pour traiter la douleur
CN104302306B (zh) 2012-03-21 2019-03-22 百深有限责任公司 Tfpi抑制剂及其使用方法
US20140099380A1 (en) 2012-06-28 2014-04-10 Ansun Biopharma, Inc. Microparticle Formulations for Delivery to the Lower and Central Respiratory Tract and Methods of Manufacture
WO2014005107A2 (fr) * 2012-06-28 2014-01-03 Ansun Biopharma, Inc. Formulations de microparticules pour la délivrance au système respiratoire supérieur et central et procédés de fabrication
CN104619369B (zh) 2012-07-12 2018-01-30 曼金德公司 干粉药物输送系统和方法
EP2911690A1 (fr) 2012-10-26 2015-09-02 MannKind Corporation Compositions et procédés de vaccin antigrippal inhalable
AU2014228415B2 (en) 2013-03-15 2018-08-09 Mannkind Corporation Microcrystalline diketopiperazine compositions and methods
CA2918369C (fr) 2013-07-18 2021-06-29 Mannkind Corporation Compositions pharmaceutiques en poudre seche stables a la chaleur et procedes
WO2015021064A1 (fr) 2013-08-05 2015-02-12 Mannkind Corporation Appareil d'insufflation et procédés
US10307464B2 (en) 2014-03-28 2019-06-04 Mannkind Corporation Use of ultrarapid acting insulin
US10561806B2 (en) 2014-10-02 2020-02-18 Mannkind Corporation Mouthpiece cover for an inhaler
US11667906B2 (en) * 2014-11-24 2023-06-06 Corning Incorporated Magnetic microcarriers
BR112018009644A2 (pt) 2015-11-12 2018-11-06 Graybug Vision Inc micropartículas agregantes sólidas modificadas na superfície, material injetável, processo para preparação de micropartículas agregantes sólidas modificadas na superfície, método para tratamento de um distúrbio ocular, e, uso de micropartículas agregantes sólidas modificadas na superfície
US20200085705A1 (en) * 2016-12-22 2020-03-19 Agency For Science, Technology And Research Method of Encapsulating Compounds
WO2018175922A1 (fr) 2017-03-23 2018-09-27 Graybug Vision, Inc. Composés et compositions pour le traitement de troubles oculaires
CN111201040A (zh) 2017-05-10 2020-05-26 灰色视觉公司 用于医学疗法的缓释微粒及其悬浮液
US11419947B2 (en) 2017-10-30 2022-08-23 Massachusetts Institute Of Technology Layer-by-layer nanoparticles for cytokine therapy in cancer treatment
WO2020115283A1 (fr) 2018-12-07 2020-06-11 Baxalta GmbH Anticorps bispécifiques fixant le facteur ixa et le facteur x
WO2020114615A1 (fr) 2018-12-07 2020-06-11 Baxalta GmbH Anticorps bispécifiques se liant au facteur ixa et au facteur x
WO2021002984A1 (fr) 2019-05-30 2021-01-07 Massachusetts Institute Of Technology Micro-aiguilles d'hydrogel fonctionnalisées par de l'acide nucléique peptidique pour l'échantillonnage et la détection d'acides nucléiques fluides interstitiels
CN114028606B (zh) * 2021-10-26 2023-06-13 湘雅生物医药(湖州)有限公司 一种壳聚糖、鱼精蛋白抗菌止血微球及其制备方法
CN114984746B (zh) * 2022-08-05 2022-11-01 山东建筑大学 一种消杀剂及养殖废水池VOCs废气处理系统方法和装置

Family Cites Families (89)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
DE2010115A1 (de) * 1970-03-04 1971-09-16 Farbenfabriken Bayer Ag, 5090 Leverkusen Verfahren zur Herstellung von Mikrogranulaten
JPS523342B2 (fr) * 1972-01-26 1977-01-27
US4197893A (en) * 1977-12-15 1980-04-15 Coin Bernard J O Reuseable puncture shield for tire casings
US4389330A (en) * 1980-10-06 1983-06-21 Stolle Research And Development Corporation Microencapsulation process
US4530840A (en) * 1982-07-29 1985-07-23 The Stolle Research And Development Corporation Injectable, long-acting microparticle formulation for the delivery of anti-inflammatory agents
JPS60100516A (ja) * 1983-11-04 1985-06-04 Takeda Chem Ind Ltd 徐放型マイクロカプセルの製造法
US4818542A (en) * 1983-11-14 1989-04-04 The University Of Kentucky Research Foundation Porous microspheres for drug delivery and methods for making same
ATE61935T1 (de) * 1985-02-07 1991-04-15 Takeda Chemical Industries Ltd Verfahren zur herstellung von mikrokapseln.
JP2551756B2 (ja) * 1985-05-07 1996-11-06 武田薬品工業株式会社 ポリオキシカルボン酸エステルおよびその製造法
US5102872A (en) * 1985-09-20 1992-04-07 Cetus Corporation Controlled-release formulations of interleukin-2
GB8601100D0 (en) * 1986-01-17 1986-02-19 Cosmas Damian Ltd Drug delivery system
EP0257915B1 (fr) * 1986-08-11 1993-03-10 Innovata Biomed Limited Formulations pharmaceutiques contenant des microcapsules
US4861627A (en) * 1987-05-01 1989-08-29 Massachusetts Institute Of Technology Preparation of multiwall polymeric microcapsules
US4897268A (en) * 1987-08-03 1990-01-30 Southern Research Institute Drug delivery system and method of making the same
US5422120A (en) * 1988-05-30 1995-06-06 Depotech Corporation Heterovesicular liposomes
USRE39497E1 (en) * 1989-02-16 2007-02-27 Nektar Therapeutics Storage of materials
AU620253B2 (en) * 1989-05-01 1992-02-13 Alkermes Controlled Therapeutics, Inc. Process for producing small particles of biologically active molecules
DK0471036T4 (da) * 1989-05-04 2004-07-19 Southern Res Inst Indkapslingsfremgangsmåde
WO1991007951A1 (fr) * 1989-12-05 1991-06-13 Trancel Corporation Composition de revêtement a base d'alginate comprenant de l'acide guluronique homologue destinee a etre appliquee et implantee in vivo et procede d'utilisation
MY107937A (en) * 1990-02-13 1996-06-29 Takeda Chemical Industries Ltd Prolonged release microcapsules.
GB9016885D0 (en) * 1990-08-01 1990-09-12 Scras Sustained release pharmaceutical compositions
US5149543A (en) * 1990-10-05 1992-09-22 Massachusetts Institute Of Technology Ionically cross-linked polymeric microcapsules
WO1992014449A1 (fr) * 1991-02-20 1992-09-03 Nova Pharmaceutical Corporation Systeme d'administration microparticulaire a liberation regulee pour des proteines
US5330768A (en) * 1991-07-05 1994-07-19 Massachusetts Institute Of Technology Controlled drug delivery using polymer/pluronic blends
US6063910A (en) * 1991-11-14 2000-05-16 The Trustees Of Princeton University Preparation of protein microparticles by supercritical fluid precipitation
US5545423A (en) * 1991-11-25 1996-08-13 Vivorx, Inc. Cytoprotective, biocompatible, retrievable macrocapsule containment systems for biologically active materials
US5525519A (en) * 1992-01-07 1996-06-11 Middlesex Sciences, Inc. Method for isolating biomolecules from a biological sample with linear polymers
US5173454A (en) * 1992-01-09 1992-12-22 Corning Incorporated Nanocrystalline materials
US5912015A (en) * 1992-03-12 1999-06-15 Alkermes Controlled Therapeutics, Inc. Modulated release from biocompatible polymers
US5286495A (en) * 1992-05-11 1994-02-15 University Of Florida Process for microencapsulating cells
US5417966A (en) * 1992-06-01 1995-05-23 Gc Corporation Depilatory composition
JP2651320B2 (ja) * 1992-07-16 1997-09-10 田辺製薬株式会社 徐放性マイクロスフェア製剤の製造方法
JP3277342B2 (ja) * 1992-09-02 2002-04-22 武田薬品工業株式会社 徐放性マイクロカプセルの製造法
EP0595030A3 (fr) * 1992-10-01 1995-06-07 Tanabe Seiyaku Co Composition de microsphères à plusieurs noyaux à libération retardée et son procédé de préparation.
EP1013270A3 (fr) * 1992-12-02 2001-03-28 Alkermes Controlled Therapeutics, Inc. Microsphères contenant une hormone de croissance à libération régulée
US5554730A (en) * 1993-03-09 1996-09-10 Middlesex Sciences, Inc. Method and kit for making a polysaccharide-protein conjugate
US6090925A (en) * 1993-03-09 2000-07-18 Epic Therapeutics, Inc. Macromolecular microparticles and methods of production and use
US5543158A (en) * 1993-07-23 1996-08-06 Massachusetts Institute Of Technology Biodegradable injectable nanoparticles
CA2176933A1 (fr) * 1993-11-18 1995-05-26 Yan Chen Preparation a liberation regulee
EP2275089A1 (fr) * 1993-11-19 2011-01-19 Alkermes Controlled Therapeutics, Inc. Préparastion de microparticules biodégradables contentant un agent biologiquement actif
US5650173A (en) * 1993-11-19 1997-07-22 Alkermes Controlled Therapeutics Inc. Ii Preparation of biodegradable microparticles containing a biologically active agent
US5620883A (en) * 1994-04-01 1997-04-15 The Johns Hopkins University Living cells microencapsulated in a polymeric membrane having two layers
US6387399B1 (en) * 1994-12-02 2002-05-14 The United States Of America As Represented By The Administrator Of The National Aeronautics And Space Administration Microencapsulated bioactive agents and method of making
US6265389B1 (en) * 1995-08-31 2001-07-24 Alkermes Controlled Therapeutics, Inc. Microencapsulation and sustained release of oligonucleotides
SE505146C2 (sv) * 1995-10-19 1997-06-30 Biogram Ab Partiklar för fördröjd frisättning
JP2909959B2 (ja) * 1995-10-19 1999-06-23 三井化学株式会社 タンパク質超薄膜固定型リアクターの製造方法及び得られたタンパク質超薄膜固定型リアクターを用いた化学反応
US5665428A (en) * 1995-10-25 1997-09-09 Macromed, Inc. Preparation of peptide containing biodegradable microspheres by melt process
US6270795B1 (en) * 1995-11-09 2001-08-07 Microbiological Research Authority Method of making microencapsulated DNA for vaccination and gene therapy
CA2192782C (fr) * 1995-12-15 2008-10-14 Nobuyuki Takechi Production de microsphere
JP2966795B2 (ja) * 1996-09-30 1999-10-25 科学技術振興事業団 機能性薄膜とその製造方法
DK0951280T3 (da) * 1996-10-03 2004-05-17 Hermes Biosciences Inc Hydrofile mikropartikler og fremgangsmåder til fremstilling heraf
US6083484A (en) * 1996-10-17 2000-07-04 Molecular Biosystems, Inc. Microparticles stabilized by polynuclear chromium complexes and their use as ultrasound contrast agents
US6395302B1 (en) * 1996-11-19 2002-05-28 Octoplus B.V. Method for the preparation of microspheres which contain colloidal systems
US5945126A (en) * 1997-02-13 1999-08-31 Oakwood Laboratories L.L.C. Continuous microsphere process
SE512663C2 (sv) * 1997-10-23 2000-04-17 Biogram Ab Inkapslingsförfarande för aktiv substans i en bionedbrytbar polymer
US6541606B2 (en) * 1997-12-31 2003-04-01 Altus Biologics Inc. Stabilized protein crystals formulations containing them and methods of making them
US6395253B2 (en) * 1998-04-23 2002-05-28 The Regents Of The University Of Michigan Microspheres containing condensed polyanionic bioactive agents and methods for their production
WO2000003797A1 (fr) * 1998-07-15 2000-01-27 MAX-PLANCK-Gesellschaft zur Förderung der Wissenschaften e.V. Revetement polyelectrolytique de specimens biologiques
US6270802B1 (en) * 1998-10-28 2001-08-07 Oakwood Laboratories L.L.C. Method and apparatus for formulating microspheres and microcapsules
DE19852928C1 (de) * 1998-11-17 2000-08-03 Steffen Panzner Strukturen in Form von Hohlkugeln
US6194006B1 (en) * 1998-12-30 2001-02-27 Alkermes Controlled Therapeutics Inc. Ii Preparation of microparticles having a selected release profile
EP1190123B1 (fr) * 1999-06-10 2002-10-16 Max-Planck-Gesellschaft zur Förderung der Wissenschaften e.V. Encapsulation de cristaux par le biais de revetements multicouches
US6458387B1 (en) * 1999-10-18 2002-10-01 Epic Therapeutics, Inc. Sustained release microspheres
DE10001172A1 (de) * 2000-01-13 2001-07-26 Max Planck Gesellschaft Templatieren von Feststoffpartikeln mit Polymermultischichten
DE10010264A1 (de) * 2000-03-02 2001-09-13 Novosom Gmbh Stabilisierte Liposomen und Hüllstrukturen
WO2001078687A1 (fr) * 2000-04-18 2001-10-25 Peptron Inc. Composition pharmaceutique injectable a liberation durable et procedes de preparation de celle-ci
AU6898001A (en) * 2000-04-28 2001-11-12 Rainer Pommersheim Method and facility for producing micromembrane capsules
FR2809309B1 (fr) * 2000-05-23 2004-06-11 Mainelab Microspheres a liberation prolongee pour administration injectable
US6998393B2 (en) * 2000-06-23 2006-02-14 Biopharm Solutions, Inc. Aquespheres, their preparation and uses thereof
KR100392501B1 (ko) * 2000-06-28 2003-07-22 동국제약 주식회사 다중 에멀젼법에 의한 서방출성 미립구의 제조방법
DE10031132A1 (de) * 2000-06-30 2002-01-17 Henkel Kgaa Verfahren zur Herstellung aktivstoffhaltiger Kapseln mit ultradünner Wandschicht
US7056554B2 (en) * 2000-08-02 2006-06-06 Max-Planck-Gesellschaft Zur Forderung Der Wissenschaften E.V. Production of polyelectrolyte capsules by surface precipitation
WO2002017888A2 (fr) * 2000-08-28 2002-03-07 MAX-PLANCK-Gesellschaft zur Förderung der Wissenschaften e.V. Gestion et support de proprietes de liberation de capsules multicouches polyelectrolytiques
US6756062B2 (en) * 2000-11-03 2004-06-29 Board Of Regents University Of Texas System Preparation of drug particles using evaporation precipitation into aqueous solutions
JP2004517699A (ja) * 2001-01-30 2004-06-17 ボード オブ リージェンツ ユニバーシティ オブ テキサス システム 液体中への噴霧凍結によるナノ粒子およびミクロ粒子の製造方法
US6896905B2 (en) * 2001-02-15 2005-05-24 Rohm And Haas Company Porous particles, their aqueous dispersions, and method of preparation
FR2824754B1 (fr) * 2001-05-15 2004-05-28 Separex Sa Procede d'obtention de particules solides a partir d'au moins un produit hydrosoluble
US6878693B2 (en) * 2001-09-28 2005-04-12 Solubest Ltd. Hydrophilic complexes of lipophilic materials and an apparatus and method for their production
AU2002334939A1 (en) * 2001-10-12 2003-04-22 Eugene R. Cooper Compositions having a combination of particles for immediate release and for controlled release
US7112361B2 (en) * 2001-10-25 2006-09-26 Massachusetts Institute Of Technology Methods of making decomposable thin films of polyelectrolytes and uses thereof
US20040014698A1 (en) * 2002-07-18 2004-01-22 Gonzalo Hortelano Oral administration of therapeutic agent coupled to transporting agent
US6896926B2 (en) * 2002-09-11 2005-05-24 Novartis Ag Method for applying an LbL coating onto a medical device
DE10244503A1 (de) * 2002-09-25 2004-04-08 Capsulution Nanoscience Ag Methode zur Herstellung und Stabilisierung von Mikro- und Nanosuspensionen mit Amphiphilen und Polyelektrolyten
AU2004280158B2 (en) * 2003-07-18 2011-02-17 Baxter International, Inc. Methods for fabrication, uses and compositions of small spherical particles prepared by controlled phase separation
US20050142205A1 (en) * 2003-07-18 2005-06-30 Julia Rashba-Step Methods for encapsulating small spherical particles prepared by controlled phase separation
EP1646354A4 (fr) * 2003-07-22 2010-03-17 Baxter Int Petites particules spheriques de molecules organiques de faible poids moleculaire et procedes de preparation et d'utilisation correspondants
WO2005112893A1 (fr) * 2004-05-12 2005-12-01 Baxter International Inc. Microsphères contenant des protéines et présentant un potentiel d’injectabilité à d’importants degrés de concentration de l’agent concerné
DK1765294T3 (da) * 2004-05-12 2008-11-10 Baxter Int Nukleinsyremikrokugler samt deres fremstilling og afgivelse
DK1758558T3 (da) * 2004-05-12 2014-01-20 Baxter Int Oligonukleotidholdige mikrokugler samt deres anvendelse til fremstilling af et medikament til behandling af diabetes type 1

Non-Patent Citations (1)

* Cited by examiner, † Cited by third party
Title
See references of WO2006116546A1 *

Also Published As

Publication number Publication date
CN101188996A (zh) 2008-05-28
WO2006116546A1 (fr) 2006-11-02
CN101188996B (zh) 2013-03-27
MX2007013356A (es) 2008-03-26
AU2006241145B2 (en) 2011-04-28
JP2008539259A (ja) 2008-11-13
AU2006241145A1 (en) 2006-11-02
US20060260777A1 (en) 2006-11-23
CA2604225A1 (fr) 2006-11-02

Similar Documents

Publication Publication Date Title
AU2006241145B2 (en) Surface-modified microparticles and methods of forming and using the same
US20100009007A1 (en) Non-covalent modification of microparticles and process of preparing same
US20080248122A1 (en) Microencapsules Containing Surface-Modified Microparticles And Methods Of Forming And Using The Same
AU2004280158B2 (en) Methods for fabrication, uses and compositions of small spherical particles prepared by controlled phase separation
KR100904931B1 (ko) 나노 입자 및 그의 제조 방법
US20070281031A1 (en) Microparticles and methods for production thereof
JP2021519748A (ja) タンパク質治療薬をカプセル製剤化および徐放性製剤化を可能にするポリマーナノ粒子組成物
EP3641737B1 (fr) Nanoparticules d'albumine pour le traitement du cancer et de maladies oculaires
CN101522177A (zh) 包含表面改性微粒的微囊及其制备和使用方法
ES2654610T3 (es) Métodos de procesamiento de composiciones que contienen micropartículas
CN109568601B (zh) 一种蛋白多肽类药物双重微球及其制备方法和胰岛素双重微球
US20100047292A1 (en) Methods of processing microparticles and compositions produced thereby
Kumar et al. Chitosan Nano Particles by Ionotropic Gelation Containing L-Arginine
RU2797114C2 (ru) Наночастица и фармацевтическая композиция для лечения глазных заболеваний или рака
Uwaezuoke et al. for Therapeutic and Diagnostic Applications: Part I: Lipids and Fabrication Techniques
CN117717524A (zh) 一种负载精氨酸的白蛋白基缓释微球及其制备方法与应用
CN120789027A (zh) 一种用于口服给药的纳米递药系统及其制备方法
Iqbal Preparation of submicron particles for theranostic applications: imaging and therapy
Samad et al. 20 Microsphere
Ganesan et al. FORMULATION AND IN VITRO EVALUATION OF CEFIXIME LIPOSPHERE
Chagar Development of injectable pH-degradable polymeric particles containing thrombin for minimally invasive delivery of fibrin glue to the heart post-myocardial infraction
HK40028224B (en) Albumin nanoparticles for the treatment of cancer and ocular diseases
FTIR Subject Index Volumes 307–325 (2006)

Legal Events

Date Code Title Description
PUAI Public reference made under article 153(3) epc to a published international application that has entered the european phase

Free format text: ORIGINAL CODE: 0009012

17P Request for examination filed

Effective date: 20071102

AK Designated contracting states

Kind code of ref document: A1

Designated state(s): AT BE BG CH CY CZ DE DK EE ES FI FR GB GR HU IE IS IT LI LT LU LV MC NL PL PT RO SE SI SK TR

DAX Request for extension of the european patent (deleted)
17Q First examination report despatched

Effective date: 20090327

RAP1 Party data changed (applicant data changed or rights of an application transferred)

Owner name: BAXTER INTERNATIONAL INC. (A DELAWARE CORPORATION)

Owner name: BAXTER HEALTHCARE S.A.

Owner name: LOUISIANA TECH UNIVERSITY

STAA Information on the status of an ep patent application or granted ep patent

Free format text: STATUS: EXAMINATION IS IN PROGRESS

STAA Information on the status of an ep patent application or granted ep patent

Free format text: STATUS: THE APPLICATION IS DEEMED TO BE WITHDRAWN

18D Application deemed to be withdrawn

Effective date: 20161101