US20080138408A1 - Drug delivery system and method - Google Patents

Drug delivery system and method Download PDF

Info

Publication number
US20080138408A1
US20080138408A1 US11/985,263 US98526307A US2008138408A1 US 20080138408 A1 US20080138408 A1 US 20080138408A1 US 98526307 A US98526307 A US 98526307A US 2008138408 A1 US2008138408 A1 US 2008138408A1
Authority
US
United States
Prior art keywords
therapeutic
drug
polymeric hydrogel
nucleic acid
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.)
Abandoned
Application number
US11/985,263
Inventor
Siddharth Venkatesh
Jacek Wower
Mark E. Byrne
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.)
Auburn University
Original Assignee
Auburn 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 Auburn University filed Critical Auburn University
Priority to US11/985,263 priority Critical patent/US20080138408A1/en
Assigned to AUBURN UNIVERSITY reassignment AUBURN UNIVERSITY ASSIGNMENT OF ASSIGNORS INTEREST (SEE DOCUMENT FOR DETAILS). Assignors: BYRNE, MARK E., VENKATESH, SIDDARTH, WOWER, JACEK
Publication of US20080138408A1 publication Critical patent/US20080138408A1/en
Priority to US14/955,309 priority patent/US10064953B2/en
Abandoned legal-status Critical Current

Links

Images

Classifications

    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K47/00—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient
    • A61K47/50—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates
    • A61K47/69—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the conjugate being characterised by physical or galenical forms, e.g. emulsion, particle, inclusion complex, stent or kit
    • A61K47/6903—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the conjugate being characterised by physical or galenical forms, e.g. emulsion, particle, inclusion complex, stent or kit the form being semi-solid, e.g. an ointment, a gel, a hydrogel or a solidifying gel
    • 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
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K47/00—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient
    • A61K47/50—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates
    • A61K47/51—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the non-active ingredient being a modifying agent
    • A61K47/54—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the non-active ingredient being a modifying agent the modifying agent being an organic compound
    • A61K47/549—Sugars, nucleosides, nucleotides or nucleic acids
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K47/00—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient
    • A61K47/50—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates
    • A61K47/51—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the non-active ingredient being a modifying agent
    • A61K47/56—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the non-active ingredient being a modifying agent the modifying agent being an organic macromolecular compound, e.g. an oligomeric, polymeric or dendrimeric molecule
    • A61K47/58—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the non-active ingredient being a modifying agent the modifying agent being an organic macromolecular compound, e.g. an oligomeric, polymeric or dendrimeric molecule obtained by reactions only involving carbon-to-carbon unsaturated bonds, e.g. poly[meth]acrylate, polyacrylamide, polystyrene, polyvinylpyrrolidone, polyvinylalcohol or polystyrene sulfonic acid resin
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K47/00—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient
    • A61K47/50—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates
    • A61K47/51—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the non-active ingredient being a modifying agent
    • A61K47/56—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the non-active ingredient being a modifying agent the modifying agent being an organic macromolecular compound, e.g. an oligomeric, polymeric or dendrimeric molecule
    • A61K47/61—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the non-active ingredient being a modifying agent the modifying agent being an organic macromolecular compound, e.g. an oligomeric, polymeric or dendrimeric molecule the organic macromolecular compound being a polysaccharide or a derivative thereof
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K47/00—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient
    • A61K47/50—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates
    • A61K47/69—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the conjugate being characterised by physical or galenical forms, e.g. emulsion, particle, inclusion complex, stent or kit
    • A61K47/6943—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the conjugate being characterised by physical or galenical forms, e.g. emulsion, particle, inclusion complex, stent or kit the form being a pill, a tablet, a lozenge or a capsule
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K47/00—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient
    • A61K47/50—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates
    • A61K47/69—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the conjugate being characterised by physical or galenical forms, e.g. emulsion, particle, inclusion complex, stent or kit
    • A61K47/6955—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the conjugate being characterised by physical or galenical forms, e.g. emulsion, particle, inclusion complex, stent or kit the form being a plaster, a bandage, a dressing or a patch
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K48/00—Medicinal preparations containing genetic material which is inserted into cells of the living body to treat genetic diseases; Gene therapy
    • A61K48/0008—Medicinal preparations containing genetic material which is inserted into cells of the living body to treat genetic diseases; Gene therapy characterised by an aspect of the 'non-active' part of the composition delivered, e.g. wherein such 'non-active' part is not delivered simultaneously with the 'active' part of the composition
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K48/00—Medicinal preparations containing genetic material which is inserted into cells of the living body to treat genetic diseases; Gene therapy
    • A61K48/0091—Purification or manufacturing processes for gene therapy compositions
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N15/00—Mutation or genetic engineering; DNA or RNA concerning genetic engineering, vectors, e.g. plasmids, or their isolation, preparation or purification; Use of hosts therefor
    • C12N15/09—Recombinant DNA-technology
    • C12N15/87—Introduction of foreign genetic material using processes not otherwise provided for, e.g. co-transformation
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K47/00—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient
    • A61K47/30—Macromolecular organic or inorganic compounds, e.g. inorganic polyphosphates
    • A61K47/34—Macromolecular compounds obtained otherwise than by reactions only involving carbon-to-carbon unsaturated bonds, e.g. polyesters, polyamino acids, polysiloxanes, polyphosphazines, copolymers of polyalkylene glycol or poloxamers

Definitions

  • the invention relates to drug delivery systems. More specifically, this invention relates to systems for and methods of controlled drug delivery using a delivery medium and nucleic acids incorporated therein.
  • Topical generally refers to, but is not limited to epicutaneous, inhalational, intranasal, vaginal, ocular surface and ear drops.
  • Enteral generally refers to, but is not limited to the digestive tract, oral cavity, gastric cavity, and rectal administration.
  • Parenteral generally refers to, but is not limited to administration by injection or infusion via intravenous, intrarterial, intramuscular, subcutaneous, transdermal, transmucosal, intradermal, intrathecal, intraosseous, intracardiac, interperitoneal, intravitreal, and inhalational.
  • RNA is emerging as an important drug target and versatile therapeutic agent because it folds into complex 3-D structures capable of expressing many enzymatic activities and because it “digitally” interferes with the flow of genetic information from DNA to proteins.
  • These novel “smart” macromolecules can be selected from random pools of RNA molecules based on their ability to bind metals, small organic compounds, nucleic acids, proteins and even entire cells and to change their inherent resistance to degradation for our benefit.
  • RNA is sensitive to degradation by ribonucleases (RNases), its stability can be easily regulated by incorporation of modified nucleotides. For example, incorporation of fluorine-CTP and -UTP (Epicentre Bio-technologies) makes RNA resistant to degradation by ubiquitous RNase A.
  • RNases ribonucleases
  • RNA molecules Another group of “smart” RNA molecules, ribozymes, are able to catalyze fundamental biological processes such as the synthesis of proteins (transpeptidation), aminoacylation of tRNA molecules (esterification), and RNA cleavage (transesterification).
  • Their discovery has changed our views of macromolecular evolution, recognizing the fact that an informational molecule can simultaneously posses enzymatic activity.
  • Ribozymes have now been described in a number of systems from bacteria through humans. The ubiquity of catalytic RNAs has prompted intensive investigation into potential applications as well as the mechanism of catalysis. The catalytic performance of nucleic acids can be enhanced by the incorporation of additional functional groups. A number of new ribozymes was discovered using SELEX.
  • the present invention is directed to methods of and system for delivering a therapeutic dose of a drug.
  • a system in accordance with the present invention includes a delivery medium with therapeutic units attached thereto.
  • the delivery medium is any suitable delivery medium that can complex or incorporate the therapeutic units.
  • the delivery medium is a gel matrix, metal particles, a polymer film, a polymer network, a metal, a polymer particle, particulate gels, particulate networks, a polymeric dendrimer, a surface conjugated with complexes or any combination thereof.
  • the therapeutic units include nucleic acid moieties with active drug portions that are controllably released to provide a therapeutic dose to a biological system or biological tissue.
  • Nucleic acid is a single or double stranded polymer or oligomer consisting of ribonucleotides, deoxyribonucleotides or their modified derivatives such as fluorinated RNA, methylated DNA, locked nucleic acids (LNA), peptide nucleic acids (PNA).
  • the Nucleic acids used in the present invention can be selected or designed. NA strands can interact to form double strands or to direct NA strands towards the molecular target using hybridization (anti-sense mechanism).
  • active drug refers broadly to a molecular species that provides a therapeutic result when administered to a biological system or tissue.
  • An active drug is, for example, an antibiotic, an anti-inflammatory, an antihistamine, an antiviral agent, a cancer drug, an anesthetic, a cycloplegic, a mydriatics, a lubricant agent, a hydrophilic agent, a decongestant, a vasoconstrictor, a vasodilater, an Immuno-suppressant, an immuno-modulating agent, an anti-glaucoma agent or a combination thereof.
  • an active drug is also in a form of strands or fragments of RNA (ribonucleic acid) and/or DNA (deoxyribonucleic acid).
  • Drug herein also refers to prodrugs, nucleic acids, glyocproteins, lipid conjugated drugs and protein-based drugs.
  • a drug herein also includes Pharmacologically active agents and can include anti-cancer drugs, analgesics, antipyretics, nonsteriodal anti-inflammatory drugs, steroidal anti-inflammatory drugs, anti-allergics, anti-histamines, anti-bacterial drugs, antibiotics, anti-infective drugs, anti-fungal drugs, statins, anti-anemia drugs, cytotoxic drugs, anti-hypertensive drugs, cholesterol lowering medications, dermatological drugs, psychotherapeutic drugs, vitamins, minerals, dietetics, anti-adiposity drugs, carbohydrate metabolism drugs, protein metabolism drugs, thyroid drugs, antithyroid drugs, anti-macular degeneration drugs, anti-retinal degenerative disease drugs, anti-diabetic retinopathydrugs, anti-uveitis drugs, anti-glaucoma drugs, immuno-modulating agents, anti-viral agents, coenzymes and combinations thereof.
  • anti-cancer drugs analgesics, antipyretics, nonsteriodal anti-inflammatory drugs,
  • a system for delivering a therapeutic dose of an active drug includes a polymeric hydrogel matrix.
  • the polymeric hydrogel matrix has therapeutic units incorporated therein.
  • the therapeutic units include strands of the NA.
  • the strands of the NA or portions thereof can act as the active drug when they are released from the polymeric hydrogel matrix.
  • the therapeutic units include strands of NA moieties.
  • NA moieties refers to strands of the NA that have drug moieties selectively bound to the stands of NA.
  • the active drug includes the drug moiety or any portion of the NA moiety that is released from the polymeric hydrogel matrix to provide a therapeutic dose.
  • the NA moieties are preferably controllably and/or selectively cleaved and/or released from polymeric hydrogel matrix to thereby release the active drug portions.
  • a drug delivery system of the present invention with different types of therapeutic units with, for example, more than one type of NA moiety and/or more than one type of drug moiety is within the scope of the preset invention.
  • a drug delivery system with different types of therapeutic units is useful to provide a controlled release of multiple active drugs in different quantities and/or at different rates.
  • a system for delivering a therapeutic dose of a drug includes metal nanoparticles.
  • the metal nanoparticles are metal particles that have an average particle size in a range of 5 to 100 nanometers in diameter.
  • the metal nanoparticles are formed from any suitable metal or combination of metals, but are preferably formed from a metal that is nontoxic or exhibits minimal toxicity to a biological system or tissue being treated.
  • the metal nanoparticles are formed from silver or gold.
  • the metal nanoparticles are functionalized with therapeutic NA moieties, such as described above, and other moieties that solubilize the metal nanoparticles and/or receptor moieties that target biological tissues or molecules.
  • the drug delivery system of the present can take any number of forms.
  • the system can be in a pill form, a patch form or a liquid form.
  • patch form includes a contact lense or any other form of a drug delivery system that administers a drug through extended contact with a biological tissue. Further details of therapeutic contact lenses are provided in the U.S. patent application Ser. No. 11/346,770, filed Feb. 3, 2006, and titled “Contact Drug Delivery System,” the contents of which are hereby incorporated by reference.
  • a drug delivery system in accordance with the present invention is taken orally, intravenously or absorbed through contact with a biological tissue.
  • the drug delivery system can be injected at or near a target tissue to be treated.
  • the polymeric hydrogel matrix with the therapeutic units provides a controlled release of an active drug.
  • the controlled release of the active drug is achieved by altering an environment of the polymeric hydrogel matrix.
  • the polymeric hydrogel matrix is heated, treated with a pH modifier, treated with an ionic solution, exposed to an electric field and/or exposed to light. In this way the release of the active drug can be stopped or started.
  • the release of the active drug is controlled over time at a rate that is regulated by a degradation rate of the NA moieties and the folding of the NA moieties in the therapeutic units.
  • a controlled release of an active drug is achieved by treating the polymeric hydrogel matrix with an enzyme that selectively cleaves the strands of the NA moieties and, thereby, releases active drug portions of the therapeutic units.
  • a drug delivery system with polymeric hydrogel matrix and therapeutic units incorporated therein is formed by providing a backbone monomer, a cross-linking monomer and NA moieties, and initiating copolymerization of the backbone monomer and cross-linking monomer.
  • the NA moieties are NA strands and/or NA strands with drug moieties complexed thereto.
  • NA strands are complexed with drug moieties prior to initiating copolymerization of the backbone monomer and cross-linking monomer, in situ with the formation of polymeric hydrogel matrix or after the polymeric hydrogel matrix is formed.
  • the backbone functional groups in the polymeric hydrogel matrix are not required to interact with the drug, but rather provide a matrix to host the drug and control its release.
  • FIG. 1A is a schematic representation showing the formation of a drug delivery system with a polymeric hydrogel matrix with therapeutic units that include NA moieties, portions of which are released by enzymatic cleavage, in accordance with the embodiments of the invention.
  • FIG. 1B illustrates a graphical representation of a controlled release profile of active drug portions of NA fragments from a drug delivery system by enzymatic cleavage within a hydrogel matrix, in accordance with the embodiments of the invention.
  • FIG. 2A is a schematic representation showing the formation of a drug delivery system with a polymeric hydrogel with therapeutic units that include strands of the NA and drug moieties complexed with strands of NA and that are released by enzymatic cleavage, in accordance with the embodiments of the invention.
  • FIG. 2B illustrates a graphical representation of a controlled release profile of drug moieties from strands of NA in a hydrogel matrix, in accordance with the embodiments of the invention.
  • FIG. 3 is a schematic representation showing the formation of a drug delivery system with a polymeric hydrogel and therapeutic units that include strands of the NA that are released by altering the environment of the polymeric hydrogel, in accordance with the embodiments of the invention.
  • FIG. 4A is a schematic representation showing the formation of a drug delivery system with metal nanoparticles that are functionalized with receptor molecules and two different therapeutic units each including NA moieties, in accordance with the embodiments of the invention.
  • FIG. 4B illustrates a graphical representation of controlled release profiles for two different active drugs released from the drug delivery system represented in FIG. 4A , in accordance with the embodiments of the invention.
  • FIG. 5 shows Structure and Function of an Intelligent Biohybrid Hydrogel with RNA Crosslinks.
  • FIGS. 6A-B show Dynamic Tunable Release Profiles of DNA from Novel Biohybrid Gels by Enzymatic or Temperature Triggers.
  • FIGS. 7A-B show Tunable Release Profiles of DNA from Biocompatible Gels by Controlling Macromolecular Architecture and Enzymatic Triggers.
  • FIG. 1A is a schematic representation 100 showing the formation of a drug delivery system 111 with a polymeric hydrogel matrix with therapeutic units 121 incorporated therein.
  • the drug delivery system 111 is formed by providing backbone monomers 101 , 103 , 105 , a cross-linking monomer (not shown), and therapeutic units 121 , and initiating copolymerization of the backbone monomers 101 , 103 , 105 and the cross-linking monomer.
  • the therapeutic units 121 are incorporated into the polymeric hydrogel matrix during the formation of the polymeric hydrogel matrix or can be incorporated into the polymeric hydrogel matrix after the polymeric hydrogel matrix is formed.
  • Polymeric hydrogel matrix of the present are incorporated into electrophoresis wells or used as membranes, films, etc. to add in the separation and capture of nucleic acid sequences. Electrophoresis and diffusion can both be used to construct a nucleic acid:drug complex within the gel or to remove unincorporated nucleic acid and drug moieties.
  • a polymeric hydrogel matrix is formed from silicon-based cross-linking monomers, carbon-based or organic-based monomers, macromers or combinations thereof.
  • Suitable cross-linking monomers include but are not limited to Polyethylene glycol (200) dimethacrylate (PEG200DMA), ethylene glycol dimethacrylate (EGDMA), tetraethyleneglycol dimethacrylate (TEGDMA), N,N′-Methylene-bis-acrylamide and polyethylene glycol (600) dimethacrylate (PEG600DMA).
  • Suitable silicon-based cross-linking monomers include, but are not limited to, tris(trimethylsiloxy)silyl propyl methacrylate (TRIS) and hydrophilic TRIS derivatives such as tris(trimethylsiloxy)silyl propyl vinyl carbamate (TPVC), tris(trimethylsiloxy)silyl propyl glycerol methacrylate (SIGMA), tris(trimethylsiloxy)silyl propyl methacryloxyethylcarbamate (TSMC); polydimethylsiloxane (PDMS) and PDMS derivatives, such as methacrylate end-capped fluoro-grafted PDMS crosslinker, a methacrylate end-capped urethane-siloxane copolymer crosslinker, a styrene-capped siloxane polymer containing polyethylene oxide and polypropylene oxide blocks; and siloxanes containing hydrophilic grafts or amino acid residue grafts
  • a solubilizing cosolvent such as dimethylsulfoxide (DMSO), isopropanol, water, alchohol, or water/alchohol mixtures.
  • DMSO dimethylsulfoxide
  • crosslinkers contain more than one vinyl group in the structure or chemistry to covalently bond with multiple monomeric or oligomeric structures.
  • Degradable crosslinkers and grafted structures degrade at a characteristic rate and include, but are not limited to poly lactic acid and polyglycolic acid macromers and derivatives, degradable thiol-ene polymers and etc.
  • Crosslinking monomer amounts from 0.01 to 90%, backbone monomers from 99.99% to 10% (moles backbone monomers/moles total monomers) with varying relative portions of backbone monomers (some of which may be functional and interact with the drug, nucleic acid, or both); initiator concentrations ranging from 0.1 to 30 wt %; solvent concentrations ranging from 0% to 80 wt %; monomers to therapeutic unit ratios ranging from 0.001 to 5,000, optimized to give a high therapeutic unit loading (therapeutic unit may be linked to network via covalent or non-covalent chemistry or contain an acrylate or methacrylate group and link to the network as other monomers in the polymerization reaction).
  • the drug delivery system 111 is formed by making a mixture or solution that includes amounts of the therapeutic units 121 , the backbone monomers 101 , 103 and 105 , the cross-linking monomer and a polymerization initiator in a suitable solvent or without a solvent.
  • Suitable initiators include water and non-water soluble initiators include, but are not limited to, TEMED (N,N,N,N-Tetramethyl-Ethylenediamine) or other reaction accelerator in conjunction with ammonium persulfate, azobisisobutyronitrile (AIBN), 2,2-dimethoxy-2-phenyl acetophenone (DMPA), 1-hydroxycyclohexyl phenyl ketone (Irgacureo 184), 2,2-dimethoxy-1,2-diphenylethan-1-one (Irgacure 651), ammonium persulfate, iniferters such as tetraethylthiuram disulfide, or combinations thereof for living or controlled polymerization methods.
  • the polymerization is able to be photo-initiated, thermally-initiated, redox-initiated or combinations thereof.
  • living or controlled polymerization examples include, but are not limited to living anionic or cationic polymerization, ring opening metathesis polymerization (ROMP), group transfer polymerization (GOP), living Ziegler-Natta polymerization, and free-radical polymerization (e.g., iniferter polymerization, catalytic chain transfer polymerization, stable free radical mediated polymerization (SFRP), ATRF or atom transfer radical polymerization, reversible addition fragmentation chain transfer (RAFT) polymerization, Iodine Transfer polymerization, Selenium-centered mediated polymerization, Telluride-mediated polymerization (TERP), Stibine-mediated polymerization).
  • SFRP stable free radical mediated polymerization
  • ATRF atom transfer radical polymerization
  • RAFT reversible addition fragmentation chain transfer
  • the backbone monomers 101 , 103 and 105 complex with the therapeutic units 121 and copolymerize with the cross-linking monomer to form the drug delivery system 111 , such as described above.
  • the monomers 101 , 103 and 105 do not complex or weakly complex with the therapeutic units 121 and copolymerize with the cross-linking monomer to form the drug delivery system 111 .
  • Functional or reactive monomers 101 and 103 useful herein are those which possess chemical or thermodynamic compatibilities with a desired therapeutic unit 121 .
  • backbone monomer includes moieties or chemical compounds that have at least one double bond group that can be incorporated into a growing polymer chain by chemical reaction and that have one end that will functionally interact with the therapeutic unit 121 through one or more of electrostatic interactions, hydrogen bonding, hydrophobic interactions, coordination complexation, and Van der Waals forces.
  • Backbone monomers include macromers, oligomers, and polymer chains with pendent functionality and which have the capability of being crosslinked to create the recognitive hydrogel.
  • Crosslinking monomers include chemicals with multiple double bond functionality that can be polymerized into a polymer network.
  • backbone monomers include, but are not limited to, 2-hydroxyethylmethacrylate (HEMA), Acrylic Acid (AA), Acrylamide (AM), N-vinyl 2-pyrrolidone (NVP), 1-vinyl-2-pyrrolidone (VP), methyl methacrylate (MMA), methacrylic acid (MAA), acetone acrylamide, 2-ethyl-2-(hydroxymethyl)-1,3-propanediol trimethacrylate, N-(1,1-dimethyl-3-oxobutyl)acrylamide, 2-ethyl-2-(hydroxymethyl)-1,3-propanediol trimethacrylate, 2,3-dihydroxypropyl methacrylate, allyl methacrylate, 3-[3,3,5,5,5-pentamethyl-1,1-bis[pentamethyldisiloxanyl)oxy]trisiloxanyl]propyl methacrylate, 3-[3,3,3-trimethyl-1,1-bis(trimethyls
  • the drug delivery system 111 is formed, it is fashioned into a pill, a contact lens, a patch or any other suitable form that allows it to be delivered to a biological system or biological tissue.
  • the system 111 is formed outside or inside the body using various methods to produce films, macroflims, microfilms, nanofilms, irregular particles and other shapes, surface coatings, particles, contact lenses with curved surfaces, etc.
  • the therapeutic units 121 preferably include NA moieties 123 , 123 ′ and 123 ′′.
  • the NA moieties include strands of NA with active drug portions 113 .
  • the drug delivery system 111 is delivered to a biological system or biological tissue using any suitable method.
  • the drug delivery system 111 is then treated with an enzyme.
  • the enzyme selectively cleaves the NA moieties 123 , 123 ′ and 123 ′′ and releases the active drug portions 113 , thereby administering a therapeutic dose of the active drug portions 113 to the biological system or biological tissue.
  • FIG. 1B illustrates a graphical representation 150 of a controlled release profile 151 of the active drug portions 113 from the drug delivery system 111 by enzymatic cleavage, in accordance with the embodiments of the invention.
  • the depleted hydrogel matrix 111 ′ FIG. 1A
  • the system is also controllable, such that a release profile 151 ′′ of the active drug portions 113 from the drug delivery system 111 is constant.
  • FIG. 2A is a schematic representation 200 showing the formation of a drug delivery system 211 , in accordance with further embodiments of the invention.
  • the drug delivery system 211 with a polymeric hydrogel matrix further includes therapeutic units 222 incorporated into the polymeric hydrogel matrix.
  • the drug delivery system 211 is formed by providing backbone monomers 201 , 203 and 205 , a cross-linking monomer (not shown) and therapeutic units 222 , and initiating copolymerization of the backbone monomers 201 , 203 and 205 and cross-linking monomer with a suitable initiator, such as described above.
  • the therapeutic units 222 preferably include NA moieties 223 .
  • the NA moieties 223 include strands of NA such as described previously.
  • the therapeutic units 222 further include drug moieties 213 that are complexed with the NA moieties 223 .
  • the drug moieties 213 are suitable drug molecules or a combination of drug molecules.
  • multiple drug moieties are used to administer therapeutic doses of the multiple drug moieties from a single drug delivery system 211 .
  • Suitable drug moieties include, but are not limited to, Anti-bacterials, Anti-infectives, Anti-microbial Agents, such as anti-fungal agents (all of which generally referred to as antibiotics) such as Penicillins (including Aminopenicillins and/or penicillinas in conjunction with penicillinase inhibitor and anti-fugal agents), Cephalosporins (and the closely related cephamycins and carbapenems), Fluoroquinolones, Tetracyclines, Macrolides, Aminoglycosides.
  • Anti-bacterials include Anti-infectives, Anti-microbial Agents, such as anti-fungal agents (all of which generally referred to as antibiotics) such as Penicillins (including Aminopenicillins and/or penicillinas in conjunction with penicillinase inhibitor and anti-fugal agents), Cephalosporins (and the closely related cephamycins and carbapenems), Fluoroquinolones, Tetracyclines, Macrolides, Aminoglycoside
  • erythromycin bacitracin zinc
  • polymyxin polymyxin B sulfates
  • neomycin gentamycin
  • tobramycin gramicidin
  • ciprofloxacin trimethoprim
  • ofloxacin levofloxacin
  • gatifloxacin moxifloxacin
  • norfloxacin sodium sulfacetamide
  • chloramphenicol tetracycline
  • azithromycin clarithyromycin, trimethoprim sulfate and bacitracin.
  • Drug delivery systems of the present invention are used to deliver Non-steroidal (NSAIDs) and Steroidal Anti-inflammatory Agents (generally referred to as anti-inflammatory agents) including both COX-1 and COX-2 inhibitors.
  • NSAIDs Non-steroidal
  • Steroidal Anti-inflammatory agents include both COX-1 and COX-2 inhibitors.
  • Examples include, but are not limited to, corticosteroids, medrysone, prednisolone, prednisolone acetate, prednisolone sodium phosphate, fluormetholone, dexamethasone, dexamethasone sodium phosphate, betamethasone, fluoromethasone, antazoline, fluorometholone acetate, rimexolone, loteprednol etabonate, diclofenac (diclofenac sodium), ketorolac, ketorolac tromethamine, hydrocortisone, bromfenac, flurbiprofen,
  • Drug delivery systems of the present invention are also used to deliver Anti-histamines, Mast cell stabilizers, and Anti-allergy Agents (generally referred to as anti-histamines).
  • anti-histamines include, but are not limited, cromolyn sodium, lodoxamide tromethamine, olopatadine HCl, nedocromil sodium, ketotifen fumurate, levocabastine HCL, azelastine HCL, pemirolast (pemirolast potassium), epinastine HCL, naphazoline HCL, emedastine, antazoline, pheniramine, sodium cromoglycate, N-acetyl-aspartyl glutamic acid and amlexanox.
  • the drug delivery systems of the present invention are used to deliver Anti-viral Agents including, but not limited to, trifluridine and vidarabine; Anti-Cancer Therapeutics including, but not limited to, dexamethasone and 5-fluorouracil (5FU); Local Anesthetics including, but are not limited to, tetracaine, proparacaine HCL and benoxinate HCL; Cycloplegics and Mydriatics including, but not limited to, Atropine sulfate, phenylephrine HCL, Cyclopentolate HCL, scopolamine HBr, homatropine HBr, tropicamide and hydroxyamphetamine Hbr; Comfort Molecules or Molecules (generally referred as lubricating agents) to treat Keratoconjunctivitis Sicca (Dry Eye) including, but not limited to, Hyaluronic acid or hyaluronan (of varying Molecular Weight, MW
  • the drug delivery system 211 is delivered to a biological system or biological tissue using any suitable method.
  • the drug delivery system 211 is then treated with an enzyme, a denaturing agent or any other suitable chemical that causes the complexed drug moieties 213 to be controllably released from the polymeric hydrogel matrix, to thereby administering a therapeutic dose of the drug moieties 213 to the biological system or biological tissue.
  • FIG. 2B illustrates a graphical representation 250 of a controlled release profile 251 of the drug moieties 213 from the drug delivery system 211 ( FIG. 2A ), in accordance with the embodiments of the invention.
  • the depleted hydrogel matrix 211 ′ preferably degrades within the biological system or biological tissue.
  • the system is also controllable, such that a release profile 251 ′′ of the active drug portions 213 from the drug delivery system 211 is constant
  • a controlled release of the active drug portions 113 ( FIG. 1A ) and/or drug moieties 213 ( FIG. 2A ) can be achieved by altering an environment of the polymeric hydrogel matrix in any number of different ways.
  • the polymeric hydrogel matrix can be heated, treated with a pH modifier, treated with an ionic solution, exposed to light to controllably release the active drug portions 113 and/or drug moieties 213 .
  • the active drug portions 113 and/or drug moieties 213 are released through bio-degradation of the NA moieties within a biological system or biological tissue.
  • controlled release of the active drug portions 113 and/or drug moieties 213 can be achieved by degradation of any portion of the systems.
  • the release of the complexed drug moieties 213 can be controlled through the affinity of the drug moieties 213 with the NA moieties 223 .
  • a strongly complexed drug moieties 213 will be released from the system 211 at a slower rate than that of a weakly complexed drug moieties 213 .
  • the drug moieties 213 will only be released from the system by degradation of the NA moieties 223 . If, however, drug moieties 213 are very weakly complexed, the drug moieties 213 will be released from the system 211 based primarily on the concentration gradient of the drug moieties 213 .
  • NA aptmer-drug complexes in accordance with the embodiment of the invention are noncovalently linked and/or dispersed within a gel structure. Release of therapeutic can be controlled by the macromolecular architecture. For instance, a tight crosslinking structure will lead to an increased transport of therapeutic from the structure but significantly deter the NA sequences depending on the size of these structures. The affinity of the NA for the therapeutic along with the structure of the gel will determine the release rate.
  • FIG. 3 is a schematic representation 300 showing the formation of a drug delivery system 303 with a polymeric hydrogel matrix 301 and therapeutic units 309 that include strands of NA.
  • the therapeutic units 309 are incorporated into the polymeric hydrogel matrix 301 after the polymeric hydrogel matrix 301 is formed.
  • the therapeutic units 309 are incorporated into the polymeric hydrogel matrix 301 by treating the polymeric hydrogel matrix 301 with the therapeutic units 309 in an ionic environment to form the drug delivery system 303 .
  • the ionic environment can, for example, be a solution of magnesium ions or any other suitable solution of metal ions.
  • the drug delivery system 303 is delivered to a biological system or biological tissue using any suitable method.
  • the drug delivery system 303 is then treated with a chelating agent, such as ethylenediamine tetraacetate, that chelates the metal ions and, thereby, releases the therapeutic units 309 from the polymeric hydrogel matrix 301 and into the biological system or biological tissue being treated.
  • a chelating agent such as ethylenediamine tetraacetate
  • FIG. 4A is a schematic representation 400 showing the formation of a drug delivery system 401 with metal nanoparticles that are functionalized with receptor molecules 431 and therapeutic units 421 .
  • the receptor molecules or ligand molecules 431 are provided to target specific cells in the body.
  • the particle surface can also be passivated using polyethylene glycol to evade immune surveillance and the reticuloendothelial system (RES) or extend circulation time (delay excretion).
  • RES reticuloendothelial system
  • the metal nanoparticles are metal particles that have an average particle size in a range of 5 to 100 nanometers in diameter and are formed from silver or gold or a combination thereof. Larger metal nanoparticles greater than 100 nanometers are considered to within the scope of the invention.
  • the therapeutic units 421 include at least two different therapeutic units 421 ′ and 421 ′′ each including NA moieties or NA moieties with drug moieties complexed thereto.
  • the different therapeutic units 421 ′ and 421 ′′ are complexed to the metal nanoparticles through any number of chemical processes.
  • the different therapeutic units 421 ′ and 421 ′′ are tailored with a thiol group that bond to a surface of the metal nanoparticles.
  • the receptor molecules 431 are provided to solubilize the metal nanoparticles and/or target specific biological tissues.
  • the drug delivery system 401 is delivered to a biological system or biological tissue using any suitable method.
  • the drug delivery system 401 or the environment of the drug delivery system 401 is then altered chemically, photo-chemically or physically, such that the different therapeutic units 421 ′ and 421 ′′ or portions of the different therapeutic units 421 ′ and 421 ′′ (active drug portions) are released from the metal nanoparticles and into the biological system or biological tissue being treated.
  • the different therapeutic units 421 ′ and 421 ′′ or portions of the different therapeutic units 421 ′ and 421 ′′ are released from the metal nanoparticles through biodegradation of the NA portions of the different therapeutic units 421 ′ and 421 ′′ within the biological environment being treated.
  • Release of the different therapeutic units 421 ′ and 421 ′′ can also occur from unfolding of the RNA, degradation of the RNA, or if the affinity of the nucleic acid-drug complex optimized-release by a concentration gradient.
  • the affinity of the nucleic acid for drug can be controlled and selected.
  • FIG. 4B illustrates a graphical representation 450 of controlled release profiles 451 and 451 ′ of two different therapeutic units 421 ′ and 421 ′′ or portions of the two different therapeutic units 421 ′ and 421 ′′ as they are released from the drug delivery system 401 represented in FIG. 4A , in accordance with the embodiments of the invention. From the controlled release profiles 451 and 451 ′ it can be seen that the different therapeutic units 421 ′ and 421 ′′ or portions of the two different therapeutic units 421 ′ and 421 ′′ are released at different rates. In this way a therapeutic dose of a drug cocktail is able to be administered to a biological system or biological tissue.
  • FIG. 5 represents a Reversible Effective Crosslinking within gel by an NA aptamer binding molecule or NA moiety 513 .
  • a conjugated or covalently attached molecule (which could be a sugar, therapeutic, biological marker, antigen, antibody, or other biologically significant moiety) to polymer chains can complex with a conjugated NA aptamer and provide reversible effective crosslinks in the gel.
  • a dispersed therapeutic 523 in the gel can then be modulated in release. When the complex is formed, transport of drug 523 is reduced and when the complex 511 is not formed the transport of drug is significantly increased leading to on-off modulated delivery.
  • the depleted polymer 511 ′ preferably biodegrades.
  • a conjugated or covalently attached molecule (which could be a sugar, therapeutic, biological marker, antigen, antibody, or other biologically significant moiety) to polymer chains can complex with a conjugated NA aptamer and provide reversible effective crosslinks in the gel.
  • a dispersed therapeutic in the gel can then be modulated in release.
  • transport of drug is reduced and when the complex is not formed or inhibited the transport of drug is significantly increased leading to on-off modulated delivery.
  • release of the dispersed therapeutic will be concentration dependent and triggered by the free molecule.
  • FIG. 6A and FIG. 6B show Dynamic Tunable Release Profiles of DNA from Novel Biohybrid Gels by Enzymatic or Temperature triggers.
  • FIG. 6A illustrates a graph 600 showing the tailored release of DNA by incubating the gels in BamHI at physiological conditions, as illustrated by the line 610 , as compared to incubating in the buffer only, as illustrated by the line 620 .
  • FIG. 6B illustrates a graph 650 showing the sigmoidal temperature dependent release of DNA, as illustrated by the line 660 .
  • FIG. 7A and FIG. 7B show Tunable Release Profiles of DNA from Biocompatible Gels by Controlling Macromolecular Architecture and Enzymatic Triggers.
  • FIG. 7A illustrates a graph 700 showing the tailored release of DNA by incubating the gels in DNase I at physiological conditions in biocompatible gels of varying crosslinking densities. Crosslinking densities, calculated on an acrylated DNA-free basis, was 0.15%, as illustrated by the line 710 , 0.22%, as illustrated by the line 711 , and 0.45%, as illustrated by the line 712 .
  • FIG. 7A illustrates a graph 700 showing the tailored release of DNA by incubating the gels in DNase I at physiological conditions in biocompatible gels of varying crosslinking densities.
  • Crosslinking densities calculated on an acrylated DNA-free basis, was 0.15%, as illustrated by the line 710 , 0.22%, as illustrated by the line 711 , and 0.45%, as illustrated by the line 712 .
  • FIG. 7B illustrates a graph 750 showing the tailored release of DNA by incubating a gel of weak crosslinking density (0.15%) in DNase I at physiological conditions, as illustrated by the line 760 , as compared to incubating in buffer only, as illustrated by the line 770 .
  • FIG. 7A highlights the release characteristics of biocompatible gels of varying crosslinking densities on treatment with a non-sequence specific endonuclease DNase I.
  • FIG. 7B conclusively shows that a DNase I trigger is responsible for the cleavage of DNA covalently incorporated within a biocompatible gel.
  • OBJECTIVE 1 The rational design, synthesis, characterization, and optimization of novel biohybrid hydrogel carriers with enhanced loading and intelligent triggered release.
  • the model therapeutic will be an anti-Human Immunodeficiency Virus type 1 therapeutic (anti-HIV deoxy-ribozyme).
  • the trigger will involve physical mechanisms (e.g., temperature, pH changes) as well as a specific biomolecular interaction, which will allow a rational tailoring and control of the therapeutic release profile.
  • the 5′ acrydite oligonucleotide and its complementary oligonucleotide are synthetically prepared and resuspended to 1 mM in Tris buffer, 0.5 mM EDTA, pH 8.
  • the complementary and non-complementary oligonucleotides are radiolabeled with ?-32P ATP using 3′phosphatase-free polynucleotide kinase and purified on a denaturing 12% polyacrylamide gel.
  • Annealing of DNA strands to form double helical DNA is confirmed by the dose-dependent addition (0, 100, 200, 300 and 500 picomoles) of acrylated oligonucleotide to an aliquot of complementary oligonucleotide (5000 cpm). Each pair of oligonucleotides are heated to 90° C. for 3 minutes and directly placed on ice. Hybridization conditions are optimized in different buffers such as water and various concentrations of Tris-HCl, pH 7.5. Samples are analyzed by electrophoresis on 12% poly(acrylamide-co-bisacrylamide) non-denaturing gel. Formation of DNA duplexes are quantified using Typhoon phosphoimager and ImageQuant software (Molecular Dynamics).
  • Digestion of the DNA helices are carried out by incubating the DNA helices with restriction enzyme and restriction enzyme buffer at 37° C. for 1 hour, as per the manufacturer's instructions. Control experiments are performed in restriction enzyme buffer only (absence of BamHI) at 37° C. for 1 hour, or on ice. Digests are analyzed by electrophoresis on non-denaturing gel. Autoradiograms are quantified using Typhoon phosphoimager and ImageQuant software.
  • Novel poly(acrylamide-co-N,N′methylenebisacrylamide-co-acrylated DNA), poly(acrylamide-co-polyethylene glycol 200 dimethacrylate-co-acrylated DNA), poly(2-hydroxyethyl methacrylate-co-polyethylene glycol 600 dimethacrylate-co-acrylated DNA) hydrogels of varying crosslinking densities are prepared via redox polymerization at 25° C. in the loading lanes of a non-denaturing gel. After polymerization, the unincorporated acrylated DNA and unhybridized 32P-labeled oligonucleotides are efficiently eluted by electrophoresis.
  • the polymer gels in the loading lanes are then cut out and submerged in 10 mM Tris buffer, pH 7.5.
  • the rest of the base gel is dried and imaged as described earlier, in order to quantify the amount of DNA incorporated into the DNA gels, and hence determine the efficiency of polymerization reaction.
  • Physiological validation of the platform Downregulation of HIV-1 Tat/Rev mRNA.
  • the physiological relevance of the platform in context of clinical medicine is demonstrated by following similar steps to release an anti-HIV deozyribozyme.
  • the double stranded helix substrate is redesigned to include a catalytic unit of a DNA enzyme that can bind to and cleave HIV-1 TAT/Rev mRNA, the coding sequence for Tat and Rev proteins, along with the BamHI recognition site.
  • Tat and Rev represent two essential proteins of human immunodeficiency virus type 1 (HIV-1), mRNAs encoding these proteins constitute frequent targets for DNA- and RNA-based gene therapy.
  • the HIV aptamer is synthesized in vitro using synthetic DNA templates under the control of promoters specific to T7 RNA polymerase. The aptamer is then radiolabeled using [5′-32P]pCp and T4 RNA ligase and purified on a 12% poly(acrylamide-co-bisacrylamide) non-denaturing gel.
  • 3′-end labeled HIV aptamer is incubated along with the DNA gels and BamHI under physiological conditions, and the cleaved RNA fragments are analyzed by electrophoresis on a denaturing gel. Autoradiograms are quantified using Typhoon phosphoimager and ImageQuant software.
  • the double stranded helix substrate includes a siRNA construct which targets and interferes with the functioning of the vascular endothelial growth factor receptor (VEGFR).
  • VEGFR vascular endothelial growth factor receptor
  • Treatment of the DNA gels with BamHI results in the cleavage of the siRNA from the gel, and downregulation of the VEGFR mRNA.
  • the VEGFR mRNA is synthesized in vitro using synthetic DNA templates under the control of promoters specific to T7 RNA polymerase.
  • the aptamer is then radiolabeled using [5′-32P]pCp and T4 RNA ligase and purified on a 12% poly(acrylamide-co-bisacrylamide) non-denaturing gel.
  • 3′-end labeled VEGFR aptamer is incubated along with the DNA gels and BamHI under physiological conditions, and the cleaved RNA fragments are analyzed by electrophoresis on a denaturing gel. Autoradiograms are quantified using Typhoon phosphoimager and ImageQuant software.
  • OBJECTIVE 2 The rational design, synthesis, characterization, and optimization of novel biohybrid hydrogel carriers which exhibit modulatory, on-off, release of a therapeutic via reversible aptamer-analyte crosslinking.
  • Biotin-binding RNA pseudoknots are synthesized in vitro using synthetic DNA templates under the control of promoters specific to T7 RNA polymerase. Large quantities of DNA are generated using the Klenow fragment of DNA Polymerase I. Integrity of the transcripts are examined by denaturing polyacrylamide gel electrophoresis. Biotin binding pseudoknots are optimized for binding efficiency using SELEX. Pseudoknots are radiolabeled using [5′-32P]pCp and T4 RNA ligase and purified on a 12% poly(acrylamide-co-bisacrylamide) non-denaturing gel.
  • Modified nucleotides are co-transcriptionally incorporated into RNA pseudoknots to render them resistant against ribonucleases and then selected for binding via affinity chromatography. For example, by modulating incorporation of fluorine-pyrimidine nucleotides (Epicentre biotechnologies) small RNAs resilient against degradation by ubiquitous RNAse A, can be produced. Protection from other RNases is achieved by posttranscriptional modification of accessible nucleotides with nucleotide specific reagents (kethoxal, DMS, DEPC).
  • Binding properties of RNA aptamers are monitored using modified Sepharoses. Columns are equilibrated with Biotin Binding Buffer (20 mM HEPES, 100 mM KCl, 15 mM MgCl2). Loading the column with samples or magnesium/EDTA buffers is followed by spinning at 1000 rpm for 20 seconds.
  • RNA based Hydrogels using Aptamer and Bioconjugate Chemistry.
  • Poly(acrylic acid-g-RNA-g-biotin) networks are prepared from poly(acrylic acid) chains of high monodispersity. Grafting of RNA and modified biotin amines to the matrix occur via commercially available cross-linking reagents (Maleimide chemistry-EDC, oxene chemistry). Hydrogels are assembled in polystyrene molds of precise dimensions.

Abstract

A system for delivering a therapeutic dose of a drug is disclosed. The system includes a delivery medium with therapeutic units attached thereto. The delivery medium is preferably a polymeric hydrogel matrix that has therapeutic units incorporated therein or metal nanoparticles with therapeutic units complexed thereto. The therapeutic units include nucleic acid moieties. The nucleic acid moieties preferably include strands of nucleic acid and drug moieties complexed with the strands of nucleic acid. Where the system includes a polymeric hydrogel matrix, an active drug is controllably released from the polymer hydrogel matrix to provide a therapeutic dose to a biological system or biological tissue. The active drug is controllably released from the hydrogel matrix by altering the environment the hydrogel matrix, or by enzymatic cleavage of the nucleic acid moieties or by a combination thereof.

Description

    RELATED APPLICATIONS
  • This application claims priority under 35 U.S.C. § 119(e) of the co-pending U.S. Provisional Application Ser. No. 60/858,553, titled “Hypersensitive Modulating Elements for Therapeutic Delivery,” filed Nov. 13, 2006 and the co-pending U.S. Provisional Application Ser. No. 60/937,773, titled “Drug Delivery Vehicles for On-Demand Therapeutic Release,” filed Jun. 28, 2007. The co-pending U.S. Provisional Application Ser. No. 60/858,553, titled “Hypersensitive Modulating Elements for Therapeutic Delivery,” filed Nov. 13, 2006 and the co-pending U.S. Provisional Application Ser. No. 60/937,773, titled “Drug Delivery Vehicles for On-Demand Therapeutic Release,” filed Jun. 28, 2007 are both hereby incorporated by reference.
    • FIELD OF THE INVENTION
  • The invention relates to drug delivery systems. More specifically, this invention relates to systems for and methods of controlled drug delivery using a delivery medium and nucleic acids incorporated therein.
    • BACKGROUND OF THE INVENTION
  • Routes of administration of drugs are commonly administered via topical, enteral (via the digestive tract), parenteral routes (injection or infusion). Topical generally refers to, but is not limited to epicutaneous, inhalational, intranasal, vaginal, ocular surface and ear drops. Enteral generally refers to, but is not limited to the digestive tract, oral cavity, gastric cavity, and rectal administration. Parenteral generally refers to, but is not limited to administration by injection or infusion via intravenous, intrarterial, intramuscular, subcutaneous, transdermal, transmucosal, intradermal, intrathecal, intraosseous, intracardiac, interperitoneal, intravitreal, and inhalational.
  • RNA is emerging as an important drug target and versatile therapeutic agent because it folds into complex 3-D structures capable of expressing many enzymatic activities and because it “digitally” interferes with the flow of genetic information from DNA to proteins. Recent studies demonstrated that RNA also constitutes an attractive material for nanotechnology because RNA molecules can be easily programmed to carry out specific functions through the incorporation of aptamers. These novel “smart” macromolecules can be selected from random pools of RNA molecules based on their ability to bind metals, small organic compounds, nucleic acids, proteins and even entire cells and to change their inherent resistance to degradation for our benefit.
  • Aptamers can be produced by a conceptually straightforward two-step process that involves in vitro synthesis of more than 1015 individual RNA molecules and screening them by column affinity chromatography. This approach is commonly known as in vitro selection or Systemic Evolution of Ligands by EXpotential enrichment (SELEX). Although RNA is sensitive to degradation by ribonucleases (RNases), its stability can be easily regulated by incorporation of modified nucleotides. For example, incorporation of fluorine-CTP and -UTP (Epicentre Bio-technologies) makes RNA resistant to degradation by ubiquitous RNase A.
  • Another group of “smart” RNA molecules, ribozymes, are able to catalyze fundamental biological processes such as the synthesis of proteins (transpeptidation), aminoacylation of tRNA molecules (esterification), and RNA cleavage (transesterification). Their discovery has changed our views of macromolecular evolution, recognizing the fact that an informational molecule can simultaneously posses enzymatic activity. Ribozymes have now been described in a number of systems from bacteria through humans. The ubiquity of catalytic RNAs has prompted intensive investigation into potential applications as well as the mechanism of catalysis. The catalytic performance of nucleic acids can be enhanced by the incorporation of additional functional groups. A number of new ribozymes was discovered using SELEX.
    • SUMMARY OF THE INVENTION
  • The present invention is directed to methods of and system for delivering a therapeutic dose of a drug. A system in accordance with the present invention includes a delivery medium with therapeutic units attached thereto. The delivery medium is any suitable delivery medium that can complex or incorporate the therapeutic units. For example, the delivery medium is a gel matrix, metal particles, a polymer film, a polymer network, a metal, a polymer particle, particulate gels, particulate networks, a polymeric dendrimer, a surface conjugated with complexes or any combination thereof. The therapeutic units include nucleic acid moieties with active drug portions that are controllably released to provide a therapeutic dose to a biological system or biological tissue.
  • Nucleic acid (NA) is a single or double stranded polymer or oligomer consisting of ribonucleotides, deoxyribonucleotides or their modified derivatives such as fluorinated RNA, methylated DNA, locked nucleic acids (LNA), peptide nucleic acids (PNA). The Nucleic acids used in the present invention can be selected or designed. NA strands can interact to form double strands or to direct NA strands towards the molecular target using hybridization (anti-sense mechanism).
  • The term “active drug” herein refers broadly to a molecular species that provides a therapeutic result when administered to a biological system or tissue. An active drug is, for example, an antibiotic, an anti-inflammatory, an antihistamine, an antiviral agent, a cancer drug, an anesthetic, a cycloplegic, a mydriatics, a lubricant agent, a hydrophilic agent, a decongestant, a vasoconstrictor, a vasodilater, an Immuno-suppressant, an immuno-modulating agent, an anti-glaucoma agent or a combination thereof. In accordance with the embodiments of the present invention, an active drug is also in a form of strands or fragments of RNA (ribonucleic acid) and/or DNA (deoxyribonucleic acid). Drug herein also refers to prodrugs, nucleic acids, glyocproteins, lipid conjugated drugs and protein-based drugs.
  • A drug herein also includes Pharmacologically active agents and can include anti-cancer drugs, analgesics, antipyretics, nonsteriodal anti-inflammatory drugs, steroidal anti-inflammatory drugs, anti-allergics, anti-histamines, anti-bacterial drugs, antibiotics, anti-infective drugs, anti-fungal drugs, statins, anti-anemia drugs, cytotoxic drugs, anti-hypertensive drugs, cholesterol lowering medications, dermatological drugs, psychotherapeutic drugs, vitamins, minerals, dietetics, anti-adiposity drugs, carbohydrate metabolism drugs, protein metabolism drugs, thyroid drugs, antithyroid drugs, anti-macular degeneration drugs, anti-retinal degenerative disease drugs, anti-diabetic retinopathydrugs, anti-uveitis drugs, anti-glaucoma drugs, immuno-modulating agents, anti-viral agents, coenzymes and combinations thereof.
  • In accordance with an embodiment of the invention, a system for delivering a therapeutic dose of an active drug includes a polymeric hydrogel matrix. The polymeric hydrogel matrix has therapeutic units incorporated therein. The therapeutic units include strands of the NA. The strands of the NA or portions thereof can act as the active drug when they are released from the polymeric hydrogel matrix. In further embodiments the therapeutic units include strands of NA moieties. As used herein, “NA moieties” refers to strands of the NA that have drug moieties selectively bound to the stands of NA. In this embodiment the active drug includes the drug moiety or any portion of the NA moiety that is released from the polymeric hydrogel matrix to provide a therapeutic dose. In use, the NA moieties are preferably controllably and/or selectively cleaved and/or released from polymeric hydrogel matrix to thereby release the active drug portions. It will be clear to one skilled in the art from the discussion below that a drug delivery system of the present invention with different types of therapeutic units with, for example, more than one type of NA moiety and/or more than one type of drug moiety is within the scope of the preset invention. A drug delivery system with different types of therapeutic units is useful to provide a controlled release of multiple active drugs in different quantities and/or at different rates.
  • In accordance with the embodiments of the invention, a system for delivering a therapeutic dose of a drug includes metal nanoparticles. The metal nanoparticles are metal particles that have an average particle size in a range of 5 to 100 nanometers in diameter. The metal nanoparticles are formed from any suitable metal or combination of metals, but are preferably formed from a metal that is nontoxic or exhibits minimal toxicity to a biological system or tissue being treated. For example, the metal nanoparticles are formed from silver or gold. The metal nanoparticles are functionalized with therapeutic NA moieties, such as described above, and other moieties that solubilize the metal nanoparticles and/or receptor moieties that target biological tissues or molecules.
  • The drug delivery system of the present can take any number of forms. For example, the system can be in a pill form, a patch form or a liquid form. As used herein “patch form” includes a contact lense or any other form of a drug delivery system that administers a drug through extended contact with a biological tissue. Further details of therapeutic contact lenses are provided in the U.S. patent application Ser. No. 11/346,770, filed Feb. 3, 2006, and titled “Contact Drug Delivery System,” the contents of which are hereby incorporated by reference.
  • In use, a drug delivery system in accordance with the present invention is taken orally, intravenously or absorbed through contact with a biological tissue. For example, the drug delivery system can be injected at or near a target tissue to be treated. Once the drug delivery system is taken, the polymeric hydrogel matrix with the therapeutic units provides a controlled release of an active drug. The controlled release of the active drug is achieved by altering an environment of the polymeric hydrogel matrix. For example, the polymeric hydrogel matrix is heated, treated with a pH modifier, treated with an ionic solution, exposed to an electric field and/or exposed to light. In this way the release of the active drug can be stopped or started. Alternatively, the release of the active drug is controlled over time at a rate that is regulated by a degradation rate of the NA moieties and the folding of the NA moieties in the therapeutic units. In a particular embodiment of the present invention, a controlled release of an active drug is achieved by treating the polymeric hydrogel matrix with an enzyme that selectively cleaves the strands of the NA moieties and, thereby, releases active drug portions of the therapeutic units.
  • In accordance with the embodiments of the present invention, a drug delivery system with polymeric hydrogel matrix and therapeutic units incorporated therein is formed by providing a backbone monomer, a cross-linking monomer and NA moieties, and initiating copolymerization of the backbone monomer and cross-linking monomer. The NA moieties are NA strands and/or NA strands with drug moieties complexed thereto. In accordance with an embodiment of the present invention NA strands are complexed with drug moieties prior to initiating copolymerization of the backbone monomer and cross-linking monomer, in situ with the formation of polymeric hydrogel matrix or after the polymeric hydrogel matrix is formed. The backbone functional groups in the polymeric hydrogel matrix are not required to interact with the drug, but rather provide a matrix to host the drug and control its release.
    • BRIEF DESCRIPTION OF THE DRAWINGS
  • FIG. 1A is a schematic representation showing the formation of a drug delivery system with a polymeric hydrogel matrix with therapeutic units that include NA moieties, portions of which are released by enzymatic cleavage, in accordance with the embodiments of the invention.
  • FIG. 1B illustrates a graphical representation of a controlled release profile of active drug portions of NA fragments from a drug delivery system by enzymatic cleavage within a hydrogel matrix, in accordance with the embodiments of the invention.
  • FIG. 2A is a schematic representation showing the formation of a drug delivery system with a polymeric hydrogel with therapeutic units that include strands of the NA and drug moieties complexed with strands of NA and that are released by enzymatic cleavage, in accordance with the embodiments of the invention.
  • FIG. 2B illustrates a graphical representation of a controlled release profile of drug moieties from strands of NA in a hydrogel matrix, in accordance with the embodiments of the invention.
  • FIG. 3 is a schematic representation showing the formation of a drug delivery system with a polymeric hydrogel and therapeutic units that include strands of the NA that are released by altering the environment of the polymeric hydrogel, in accordance with the embodiments of the invention.
  • FIG. 4A is a schematic representation showing the formation of a drug delivery system with metal nanoparticles that are functionalized with receptor molecules and two different therapeutic units each including NA moieties, in accordance with the embodiments of the invention.
  • FIG. 4B illustrates a graphical representation of controlled release profiles for two different active drugs released from the drug delivery system represented in FIG. 4A, in accordance with the embodiments of the invention.
  • FIG. 5 shows Structure and Function of an Intelligent Biohybrid Hydrogel with RNA Crosslinks.
  • FIGS. 6A-B show Dynamic Tunable Release Profiles of DNA from Novel Biohybrid Gels by Enzymatic or Temperature Triggers.
  • FIGS. 7A-B show Tunable Release Profiles of DNA from Biocompatible Gels by Controlling Macromolecular Architecture and Enzymatic Triggers.
    •  DETAILED DESCRIPTION OF A PREFERRED EMBODIMENT
  • FIG. 1A is a schematic representation 100 showing the formation of a drug delivery system 111 with a polymeric hydrogel matrix with therapeutic units 121 incorporated therein. The drug delivery system 111 is formed by providing backbone monomers 101, 103, 105, a cross-linking monomer (not shown), and therapeutic units 121, and initiating copolymerization of the backbone monomers 101, 103, 105 and the cross-linking monomer. The therapeutic units 121 are incorporated into the polymeric hydrogel matrix during the formation of the polymeric hydrogel matrix or can be incorporated into the polymeric hydrogel matrix after the polymeric hydrogel matrix is formed.
  • Polymeric hydrogel matrix of the present are incorporated into electrophoresis wells or used as membranes, films, etc. to add in the separation and capture of nucleic acid sequences. Electrophoresis and diffusion can both be used to construct a nucleic acid:drug complex within the gel or to remove unincorporated nucleic acid and drug moieties.
  • In accordance with the embodiments of the invention, a polymeric hydrogel matrix is formed from silicon-based cross-linking monomers, carbon-based or organic-based monomers, macromers or combinations thereof. Suitable cross-linking monomers include but are not limited to Polyethylene glycol (200) dimethacrylate (PEG200DMA), ethylene glycol dimethacrylate (EGDMA), tetraethyleneglycol dimethacrylate (TEGDMA), N,N′-Methylene-bis-acrylamide and polyethylene glycol (600) dimethacrylate (PEG600DMA). Suitable silicon-based cross-linking monomers include, but are not limited to, tris(trimethylsiloxy)silyl propyl methacrylate (TRIS) and hydrophilic TRIS derivatives such as tris(trimethylsiloxy)silyl propyl vinyl carbamate (TPVC), tris(trimethylsiloxy)silyl propyl glycerol methacrylate (SIGMA), tris(trimethylsiloxy)silyl propyl methacryloxyethylcarbamate (TSMC); polydimethylsiloxane (PDMS) and PDMS derivatives, such as methacrylate end-capped fluoro-grafted PDMS crosslinker, a methacrylate end-capped urethane-siloxane copolymer crosslinker, a styrene-capped siloxane polymer containing polyethylene oxide and polypropylene oxide blocks; and siloxanes containing hydrophilic grafts or amino acid residue grafts, and siloxanes containing hydrophilic blocks or containing amino acid residue grafts. The molecular structure of these monomers can be altered chemically to contain moieties that match amino acid residues or other biological molecules. In cases where the above monomers are polymerized with hydrophilic monomers, a solubilizing cosolvent may be used such as dimethylsulfoxide (DMSO), isopropanol, water, alchohol, or water/alchohol mixtures.
  • Preferably, crosslinkers contain more than one vinyl group in the structure or chemistry to covalently bond with multiple monomeric or oligomeric structures. Degradable crosslinkers and grafted structures degrade at a characteristic rate and include, but are not limited to poly lactic acid and polyglycolic acid macromers and derivatives, degradable thiol-ene polymers and etc.
  • Crosslinking monomer amounts from 0.01 to 90%, backbone monomers from 99.99% to 10% (moles backbone monomers/moles total monomers) with varying relative portions of backbone monomers (some of which may be functional and interact with the drug, nucleic acid, or both); initiator concentrations ranging from 0.1 to 30 wt %; solvent concentrations ranging from 0% to 80 wt %; monomers to therapeutic unit ratios ranging from 0.001 to 5,000, optimized to give a high therapeutic unit loading (therapeutic unit may be linked to network via covalent or non-covalent chemistry or contain an acrylate or methacrylate group and link to the network as other monomers in the polymerization reaction).
  • Still referring to FIG. 1, in accordance with a method of the present invention, the drug delivery system 111 is formed by making a mixture or solution that includes amounts of the therapeutic units 121, the backbone monomers 101, 103 and 105, the cross-linking monomer and a polymerization initiator in a suitable solvent or without a solvent. Suitable initiators include water and non-water soluble initiators include, but are not limited to, TEMED (N,N,N,N-Tetramethyl-Ethylenediamine) or other reaction accelerator in conjunction with ammonium persulfate, azobisisobutyronitrile (AIBN), 2,2-dimethoxy-2-phenyl acetophenone (DMPA), 1-hydroxycyclohexyl phenyl ketone (Irgacureo 184), 2,2-dimethoxy-1,2-diphenylethan-1-one (Irgacure 651), ammonium persulfate, iniferters such as tetraethylthiuram disulfide, or combinations thereof for living or controlled polymerization methods. The polymerization is able to be photo-initiated, thermally-initiated, redox-initiated or combinations thereof.
  • Examples of living or controlled polymerization include, but are not limited to living anionic or cationic polymerization, ring opening metathesis polymerization (ROMP), group transfer polymerization (GOP), living Ziegler-Natta polymerization, and free-radical polymerization (e.g., iniferter polymerization, catalytic chain transfer polymerization, stable free radical mediated polymerization (SFRP), ATRF or atom transfer radical polymerization, reversible addition fragmentation chain transfer (RAFT) polymerization, Iodine Transfer polymerization, Selenium-centered mediated polymerization, Telluride-mediated polymerization (TERP), Stibine-mediated polymerization).
  • The backbone monomers 101, 103 and 105 complex with the therapeutic units 121 and copolymerize with the cross-linking monomer to form the drug delivery system 111, such as described above. Alternatively, the monomers 101, 103 and 105 do not complex or weakly complex with the therapeutic units 121 and copolymerize with the cross-linking monomer to form the drug delivery system 111. Functional or reactive monomers 101 and 103 useful herein are those which possess chemical or thermodynamic compatibilities with a desired therapeutic unit 121. As used herein, the term backbone monomer includes moieties or chemical compounds that have at least one double bond group that can be incorporated into a growing polymer chain by chemical reaction and that have one end that will functionally interact with the therapeutic unit 121 through one or more of electrostatic interactions, hydrogen bonding, hydrophobic interactions, coordination complexation, and Van der Waals forces. Backbone monomers include macromers, oligomers, and polymer chains with pendent functionality and which have the capability of being crosslinked to create the recognitive hydrogel. Crosslinking monomers include chemicals with multiple double bond functionality that can be polymerized into a polymer network.
  • Examples of backbone monomers include, but are not limited to, 2-hydroxyethylmethacrylate (HEMA), Acrylic Acid (AA), Acrylamide (AM), N-vinyl 2-pyrrolidone (NVP), 1-vinyl-2-pyrrolidone (VP), methyl methacrylate (MMA), methacrylic acid (MAA), acetone acrylamide, 2-ethyl-2-(hydroxymethyl)-1,3-propanediol trimethacrylate, N-(1,1-dimethyl-3-oxobutyl)acrylamide, 2-ethyl-2-(hydroxymethyl)-1,3-propanediol trimethacrylate, 2,3-dihydroxypropyl methacrylate, allyl methacrylate, 3-[3,3,5,5,5-pentamethyl-1,1-bis[pentamethyldisiloxanyl)oxy]trisiloxanyl]propyl methacrylate, 3-[3,3,3-trimethyl-1,1-bis(trimethylsiloxy)disiloxanyl]propyl methacrylate (TRIS), N-(1,1-dimethyl-3-oxybutyl)acrylamide, dimethyl itaconate, 2,2,2,-trifluoro-1-(trifluoromethyl)ethyl methacrylate, 2,2,2-trifluoroethyl methacrylate, methacryloxypropylbis(trimethylsiloxy)methylsilane, methacryloxypropylpentamethyldisiloxane, (3-methacryloxy-2-hydroxypropyloxy)propylbis(trimethylsiloxy)methylsilane, 4-t-butyl-2-hydroxycyclohexyl methacrylate, dimethylacrylamide, glycerol methacrylate and diethylaminoethyl methacrylate (DEAEM).
  • Once the drug delivery system 111 is formed, it is fashioned into a pill, a contact lens, a patch or any other suitable form that allows it to be delivered to a biological system or biological tissue. The system 111 is formed outside or inside the body using various methods to produce films, macroflims, microfilms, nanofilms, irregular particles and other shapes, surface coatings, particles, contact lenses with curved surfaces, etc.
  • Still referring to FIG. 1A, the therapeutic units 121 preferably include NA moieties 123, 123′ and 123″. The NA moieties include strands of NA with active drug portions 113. In use the drug delivery system 111 is delivered to a biological system or biological tissue using any suitable method. The drug delivery system 111 is then treated with an enzyme. The enzyme selectively cleaves the NA moieties 123, 123′ and 123″ and releases the active drug portions 113, thereby administering a therapeutic dose of the active drug portions 113 to the biological system or biological tissue.
  • FIG. 1B illustrates a graphical representation 150 of a controlled release profile 151 of the active drug portions 113 from the drug delivery system 111 by enzymatic cleavage, in accordance with the embodiments of the invention. After the therapeutic dose of the active drug portion 113 is released to the biological system or biological tissue, the depleted hydrogel matrix 111′ (FIG. 1A) preferably degrades within the biological system or biological tissue. The system is also controllable, such that a release profile 151″ of the active drug portions 113 from the drug delivery system 111 is constant.
  • FIG. 2A is a schematic representation 200 showing the formation of a drug delivery system 211, in accordance with further embodiments of the invention. The drug delivery system 211 with a polymeric hydrogel matrix further includes therapeutic units 222 incorporated into the polymeric hydrogel matrix. The drug delivery system 211 is formed by providing backbone monomers 201, 203 and 205, a cross-linking monomer (not shown) and therapeutic units 222, and initiating copolymerization of the backbone monomers 201, 203 and 205 and cross-linking monomer with a suitable initiator, such as described above.
  • Still referring to FIG. 2A, the therapeutic units 222 preferably include NA moieties 223. The NA moieties 223 include strands of NA such as described previously. The therapeutic units 222 further include drug moieties 213 that are complexed with the NA moieties 223. The drug moieties 213 are suitable drug molecules or a combination of drug molecules. In accordance with the embodiments of the invention, multiple drug moieties are used to administer therapeutic doses of the multiple drug moieties from a single drug delivery system 211.
  • Suitable drug moieties include, but are not limited to, Anti-bacterials, Anti-infectives, Anti-microbial Agents, such as anti-fungal agents (all of which generally referred to as antibiotics) such as Penicillins (including Aminopenicillins and/or penicillinas in conjunction with penicillinase inhibitor and anti-fugal agents), Cephalosporins (and the closely related cephamycins and carbapenems), Fluoroquinolones, Tetracyclines, Macrolides, Aminoglycosides. Specific examples include, but are not limited to, erythromycin, bacitracin zinc, polymyxin, polymyxin B sulfates, neomycin, gentamycin, tobramycin, gramicidin, ciprofloxacin, trimethoprim, ofloxacin, levofloxacin, gatifloxacin, moxifloxacin, norfloxacin, sodium sulfacetamide, chloramphenicol, tetracycline, azithromycin, clarithyromycin, trimethoprim sulfate and bacitracin.
  • Drug delivery systems of the present invention are used to deliver Non-steroidal (NSAIDs) and Steroidal Anti-inflammatory Agents (generally referred to as anti-inflammatory agents) including both COX-1 and COX-2 inhibitors. Examples include, but are not limited to, corticosteroids, medrysone, prednisolone, prednisolone acetate, prednisolone sodium phosphate, fluormetholone, dexamethasone, dexamethasone sodium phosphate, betamethasone, fluoromethasone, antazoline, fluorometholone acetate, rimexolone, loteprednol etabonate, diclofenac (diclofenac sodium), ketorolac, ketorolac tromethamine, hydrocortisone, bromfenac, flurbiprofen, antazoline and xylometazoline.
  • Drug delivery systems of the present invention are also used to deliver Anti-histamines, Mast cell stabilizers, and Anti-allergy Agents (generally referred to as anti-histamines). Examples include, but are not limited, cromolyn sodium, lodoxamide tromethamine, olopatadine HCl, nedocromil sodium, ketotifen fumurate, levocabastine HCL, azelastine HCL, pemirolast (pemirolast potassium), epinastine HCL, naphazoline HCL, emedastine, antazoline, pheniramine, sodium cromoglycate, N-acetyl-aspartyl glutamic acid and amlexanox.
  • In yet further embodiments of the invention, the drug delivery systems of the present invention are used to deliver Anti-viral Agents including, but not limited to, trifluridine and vidarabine; Anti-Cancer Therapeutics including, but not limited to, dexamethasone and 5-fluorouracil (5FU); Local Anesthetics including, but are not limited to, tetracaine, proparacaine HCL and benoxinate HCL; Cycloplegics and Mydriatics including, but not limited to, Atropine sulfate, phenylephrine HCL, Cyclopentolate HCL, scopolamine HBr, homatropine HBr, tropicamide and hydroxyamphetamine Hbr; Comfort Molecules or Molecules (generally referred as lubricating agents) to treat Keratoconjunctivitis Sicca (Dry Eye) including, but not limited to, Hyaluronic acid or hyaluronan (of varying Molecular Weight, MW), hydroxypropyl cellulose (of varying MW), gefarnate, hydroxyeicosatetranenoic acid (15-(S)-HETE), phospholipid-HETE derivatives, phoshoroylcholine or other polar lipids, carboxymethyl cellulose (of varying MW), polyethylene glycol (of varying MW), polyvinyl alcohol (of varying MW), rebamipide, pimecrolimus, ecabet sodium and hydrophilic polymers; Immuno-suppressive and Immuno-modulating Agents including, but not limited to, Cyclosporine, tacrolimus, anti-IgE and cytokine antagonists; and Anti-Glaucoma Agents including beta blockers, pilocarpine, direct-acting miotics, prostagladins, alpha adrenergic agonists, carbonic anhydrase inhibitors including, but not limited to betaxolol HCL, levobunolol HCL, metipranolol HCL, timolol maleate or hemihydrate, carteolol HCL, carbachol, pilocarpine HCL, latanoprost, bimatoprost, travoprost, brimonidine tartrate, apraclonidine HCL, brinzolamide and dorzolamide HCL; decongestants, vasodilaters vasoconstrictors including, but not limited to epinephrine and pseudoephedrine.
  • Still referring to FIG. 2A, in use, the drug delivery system 211 is delivered to a biological system or biological tissue using any suitable method. The drug delivery system 211 is then treated with an enzyme, a denaturing agent or any other suitable chemical that causes the complexed drug moieties 213 to be controllably released from the polymeric hydrogel matrix, to thereby administering a therapeutic dose of the drug moieties 213 to the biological system or biological tissue.
  • FIG. 2B illustrates a graphical representation 250 of a controlled release profile 251 of the drug moieties 213 from the drug delivery system 211 (FIG. 2A), in accordance with the embodiments of the invention. After the therapeutic dose of the drug moieties 213 is released to the biological system or biological tissue, the depleted hydrogel matrix 211′ preferably degrades within the biological system or biological tissue. The system is also controllable, such that a release profile 251″ of the active drug portions 213 from the drug delivery system 211 is constant
  • It will be clear to one skilled in the art that a controlled release of the active drug portions 113 (FIG. 1A) and/or drug moieties 213 (FIG. 2A) can be achieved by altering an environment of the polymeric hydrogel matrix in any number of different ways. For example, the polymeric hydrogel matrix can be heated, treated with a pH modifier, treated with an ionic solution, exposed to light to controllably release the active drug portions 113 and/or drug moieties 213. Alternatively, the active drug portions 113 and/or drug moieties 213 are released through bio-degradation of the NA moieties within a biological system or biological tissue. Also and described previously, controlled release of the active drug portions 113 and/or drug moieties 213 can be achieved by degradation of any portion of the systems.
  • The release of the complexed drug moieties 213 (FIG. 2A) can be controlled through the affinity of the drug moieties 213 with the NA moieties 223. For example, a strongly complexed drug moieties 213 will be released from the system 211 at a slower rate than that of a weakly complexed drug moieties 213. In some cases where drug moieties 213 are very strongly complexed, the drug moieties 213 will only be released from the system by degradation of the NA moieties 223. If, however, drug moieties 213 are very weakly complexed, the drug moieties 213 will be released from the system 211 based primarily on the concentration gradient of the drug moieties 213.
  • Also, NA aptmer-drug complexes in accordance with the embodiment of the invention are noncovalently linked and/or dispersed within a gel structure. Release of therapeutic can be controlled by the macromolecular architecture. For instance, a tight crosslinking structure will lead to an increased transport of therapeutic from the structure but significantly deter the NA sequences depending on the size of these structures. The affinity of the NA for the therapeutic along with the structure of the gel will determine the release rate.
  • FIG. 3 is a schematic representation 300 showing the formation of a drug delivery system 303 with a polymeric hydrogel matrix 301 and therapeutic units 309 that include strands of NA. The therapeutic units 309 are incorporated into the polymeric hydrogel matrix 301 after the polymeric hydrogel matrix 301 is formed. The therapeutic units 309 are incorporated into the polymeric hydrogel matrix 301 by treating the polymeric hydrogel matrix 301 with the therapeutic units 309 in an ionic environment to form the drug delivery system 303. The ionic environment can, for example, be a solution of magnesium ions or any other suitable solution of metal ions.
  • Still referring to FIG. 3, in use, the drug delivery system 303 is delivered to a biological system or biological tissue using any suitable method. The drug delivery system 303 is then treated with a chelating agent, such as ethylenediamine tetraacetate, that chelates the metal ions and, thereby, releases the therapeutic units 309 from the polymeric hydrogel matrix 301 and into the biological system or biological tissue being treated.
  • FIG. 4A is a schematic representation 400 showing the formation of a drug delivery system 401 with metal nanoparticles that are functionalized with receptor molecules 431 and therapeutic units 421. The receptor molecules or ligand molecules 431 are provided to target specific cells in the body. The particle surface can also be passivated using polyethylene glycol to evade immune surveillance and the reticuloendothelial system (RES) or extend circulation time (delay excretion).
  • The metal nanoparticles are metal particles that have an average particle size in a range of 5 to 100 nanometers in diameter and are formed from silver or gold or a combination thereof. Larger metal nanoparticles greater than 100 nanometers are considered to within the scope of the invention.
  • In accordance with the embodiments of this invention, the therapeutic units 421 include at least two different therapeutic units 421′ and 421″ each including NA moieties or NA moieties with drug moieties complexed thereto. The different therapeutic units 421′ and 421″ are complexed to the metal nanoparticles through any number of chemical processes. For example the different therapeutic units 421′ and 421″ are tailored with a thiol group that bond to a surface of the metal nanoparticles. The receptor molecules 431 are provided to solubilize the metal nanoparticles and/or target specific biological tissues.
  • Still referring to FIG. 4A, in use, the drug delivery system 401 is delivered to a biological system or biological tissue using any suitable method. The drug delivery system 401 or the environment of the drug delivery system 401 is then altered chemically, photo-chemically or physically, such that the different therapeutic units 421′ and 421″ or portions of the different therapeutic units 421′ and 421″ (active drug portions) are released from the metal nanoparticles and into the biological system or biological tissue being treated. Alternatively, the different therapeutic units 421′ and 421″ or portions of the different therapeutic units 421′ and 421″ are released from the metal nanoparticles through biodegradation of the NA portions of the different therapeutic units 421′ and 421″ within the biological environment being treated. Release of the different therapeutic units 421′ and 421″ can also occur from unfolding of the RNA, degradation of the RNA, or if the affinity of the nucleic acid-drug complex optimized-release by a concentration gradient. The affinity of the nucleic acid for drug can be controlled and selected.
  • FIG. 4B illustrates a graphical representation 450 of controlled release profiles 451 and 451′ of two different therapeutic units 421′ and 421″ or portions of the two different therapeutic units 421′ and 421″ as they are released from the drug delivery system 401 represented in FIG. 4A, in accordance with the embodiments of the invention. From the controlled release profiles 451 and 451′ it can be seen that the different therapeutic units 421′ and 421″ or portions of the two different therapeutic units 421′ and 421″ are released at different rates. In this way a therapeutic dose of a drug cocktail is able to be administered to a biological system or biological tissue.
  • FIG. 5 represents a Reversible Effective Crosslinking within gel by an NA aptamer binding molecule or NA moiety 513. A conjugated or covalently attached molecule (which could be a sugar, therapeutic, biological marker, antigen, antibody, or other biologically significant moiety) to polymer chains can complex with a conjugated NA aptamer and provide reversible effective crosslinks in the gel. A dispersed therapeutic 523 in the gel can then be modulated in release. When the complex is formed, transport of drug 523 is reduced and when the complex 511 is not formed the transport of drug is significantly increased leading to on-off modulated delivery. Also, if a free molecule enters the gel that is similar to or the same as the covalently attached molecule, release of the dispersed therapeutic will be concentration dependent and triggered by the free molecule. After the drug 523 is released, the depleted polymer 511′ preferably biodegrades.
  • A conjugated or covalently attached molecule (which could be a sugar, therapeutic, biological marker, antigen, antibody, or other biologically significant moiety) to polymer chains can complex with a conjugated NA aptamer and provide reversible effective crosslinks in the gel. A dispersed therapeutic in the gel can then be modulated in release. When the complex is formed transport of drug is reduced and when the complex is not formed or inhibited the transport of drug is significantly increased leading to on-off modulated delivery. Also, if a free molecule enters the gel that is similar to or the same as the covalently attached molecule, release of the dispersed therapeutic will be concentration dependent and triggered by the free molecule.
  • Despite entrapment in the network, the DNA helix can be readily cleaved by restriction enzyme, leading to controlled release of the cleaved DNA strands (FIG. 6A). FIG. 6A and FIG. 6B show Dynamic Tunable Release Profiles of DNA from Novel Biohybrid Gels by Enzymatic or Temperature triggers. FIG. 6A illustrates a graph 600 showing the tailored release of DNA by incubating the gels in BamHI at physiological conditions, as illustrated by the line 610, as compared to incubating in the buffer only, as illustrated by the line 620. FIG. 6B illustrates a graph 650 showing the sigmoidal temperature dependent release of DNA, as illustrated by the line 660. FIG. 6B highlights the temperature responsive release characteristics which match the theoretical melting temperature (58° C.). Since melting temperature is a direct function of G-C pairs, this will provide the opportunity to tune the release characteristics by varying the sequence. Optimization of the macromolecular structure of the network or the DNA molecules will lead to programmable release profiles.
  • FIG. 7A and FIG. 7B show Tunable Release Profiles of DNA from Biocompatible Gels by Controlling Macromolecular Architecture and Enzymatic Triggers. FIG. 7A illustrates a graph 700 showing the tailored release of DNA by incubating the gels in DNase I at physiological conditions in biocompatible gels of varying crosslinking densities. Crosslinking densities, calculated on an acrylated DNA-free basis, was 0.15%, as illustrated by the line 710, 0.22%, as illustrated by the line 711, and 0.45%, as illustrated by the line 712. FIG. 7B illustrates a graph 750 showing the tailored release of DNA by incubating a gel of weak crosslinking density (0.15%) in DNase I at physiological conditions, as illustrated by the line 760, as compared to incubating in buffer only, as illustrated by the line 770. FIG. 7A highlights the release characteristics of biocompatible gels of varying crosslinking densities on treatment with a non-sequence specific endonuclease DNase I. FIG. 7B conclusively shows that a DNase I trigger is responsible for the cleavage of DNA covalently incorporated within a biocompatible gel.
    • Experimental Section
  • OBJECTIVE 1: The rational design, synthesis, characterization, and optimization of novel biohybrid hydrogel carriers with enhanced loading and intelligent triggered release. The model therapeutic will be an anti-Human Immunodeficiency Virus type 1 therapeutic (anti-HIV deoxy-ribozyme). The trigger will involve physical mechanisms (e.g., temperature, pH changes) as well as a specific biomolecular interaction, which will allow a rational tailoring and control of the therapeutic release profile.
    • Specific Aims: 1) Analyze the critical factors for hybridization and restriction enzyme digestion of the acrylated DNA helix; 2) Synthesize novel hydrogels with a covalently linked pendant DNA helix via redox polymerization and within the gel; 3) Characterize and optimize loading efficiency of acrylated DNA helix; 4) Analyze the in-vitro dynamic release of the cleaved DNA from the network via temperature change, pH change, a sequence specific enzymatic reaction; 5) Conduct diffusional analysis of cleaved DNA and restriction enzyme through network; 6) Synthesize networks of varying macromolecular structures (e.g., crosslinking densities) and determine the release characteristics of the cleaved DNA from biocompatible gels (e.g., synthesized from FDA-approved monomers); 7) Demonstrate physiological relevance of system with dynamic release studies of anti-HIV deoxyribozyme; 8) Perform network structural analysis via equilibrium weight and volume swelling studies. Experiments:
  • In-vitro Hybridization. The 5′ acrydite oligonucleotide and its complementary oligonucleotide are synthetically prepared and resuspended to 1 mM in Tris buffer, 0.5 mM EDTA, pH 8. The complementary and non-complementary oligonucleotides are radiolabeled with ?-32P ATP using 3′phosphatase-free polynucleotide kinase and purified on a denaturing 12% polyacrylamide gel. Annealing of DNA strands to form double helical DNA is confirmed by the dose-dependent addition (0, 100, 200, 300 and 500 picomoles) of acrylated oligonucleotide to an aliquot of complementary oligonucleotide (5000 cpm). Each pair of oligonucleotides are heated to 90° C. for 3 minutes and directly placed on ice. Hybridization conditions are optimized in different buffers such as water and various concentrations of Tris-HCl, pH 7.5. Samples are analyzed by electrophoresis on 12% poly(acrylamide-co-bisacrylamide) non-denaturing gel. Formation of DNA duplexes are quantified using Typhoon phosphoimager and ImageQuant software (Molecular Dynamics).
  • In-vitro Restriction Enzyme Digestion. Digestion of the DNA helices are carried out by incubating the DNA helices with restriction enzyme and restriction enzyme buffer at 37° C. for 1 hour, as per the manufacturer's instructions. Control experiments are performed in restriction enzyme buffer only (absence of BamHI) at 37° C. for 1 hour, or on ice. Digests are analyzed by electrophoresis on non-denaturing gel. Autoradiograms are quantified using Typhoon phosphoimager and ImageQuant software.
  • Synthesis and Characterization of Biocompatible DNA Gels. Novel poly(acrylamide-co-N,N′methylenebisacrylamide-co-acrylated DNA), poly(acrylamide-co-polyethylene glycol 200 dimethacrylate-co-acrylated DNA), poly(2-hydroxyethyl methacrylate-co-polyethylene glycol 600 dimethacrylate-co-acrylated DNA) hydrogels of varying crosslinking densities are prepared via redox polymerization at 25° C. in the loading lanes of a non-denaturing gel. After polymerization, the unincorporated acrylated DNA and unhybridized 32P-labeled oligonucleotides are efficiently eluted by electrophoresis. The polymer gels in the loading lanes are then cut out and submerged in 10 mM Tris buffer, pH 7.5. The rest of the base gel is dried and imaged as described earlier, in order to quantify the amount of DNA incorporated into the DNA gels, and hence determine the efficiency of polymerization reaction.
  • In-vitro Kinetic Release of DNA Strands upon Restriction Enzyme Trigger. Release studies of 32P-labeled loaded DNA are conducted by incubating the DNA gels under physiological conditions in the presence of BamHI and BamHI buffer, and the release is monitored by Cerenkov counting. Control experiments are performed, by incubating the DNA gels in BamHI buffer (absence of BamHI), and by using the restriction endonuclease EcoRI, which does not recognize the BamHI recognition site. The enzymatic activity of BamHI is lowered by decreasing the pH and release will be monitored. Each experiment is performed five times and cumulative and differential DNA released versus time, normalized dynamic release profiles, and drug diffusion coefficient will be calculated by using Fick's Law with a diffusion coefficient that is dependent on position via one-dimensional planar solute release from the gel.
  • In-vitro Kinetic Release of DNA Strands upon Deoxynuclease I Trigger. Release studies of 32P-labeled loaded DNA are conducted by incubating the DNA gels under physiological conditions in the presence of DNase I (which is a non-specific endonuclease), and the release is monitored by Cerenkov counting. Control experiments are performed, by incubating the DNA gels in DNase I buffer (absence of DNaseI). The effect of the macromolecular architecture the diffusion of DNA strands from the gel to give tunable release profiles are investigated by conducting release studies in gels of varying crosslinking densities. Each experiment is performed five times and cumulative and differential DNA released versus time, normalized dynamic release profiles, and drug diffusion coefficient will be calculated by using Fick's Law with a diffusion coefficient that is dependent on position via one-dimensional planar solute release from the gel.
  • In-vitro Kinetic Release of DNA Strands upon Temperature Ramp Temperature-responsive release of melted DNA strands are investigated between 30° C. and 85° C. Each experiment is performed five times and cumulative and differential DNA released versus time, normalized dynamic release profiles, and drug diffusion coefficient will be calculated by using Fick's Law with a diffusion coefficient that is dependent on position via one-dimensional planar solute release from the gel.
  • Physiological validation of the platform: Downregulation of HIV-1 Tat/Rev mRNA. The physiological relevance of the platform in context of clinical medicine is demonstrated by following similar steps to release an anti-HIV deozyribozyme. The double stranded helix substrate is redesigned to include a catalytic unit of a DNA enzyme that can bind to and cleave HIV-1 TAT/Rev mRNA, the coding sequence for Tat and Rev proteins, along with the BamHI recognition site. As Tat and Rev represent two essential proteins of human immunodeficiency virus type 1 (HIV-1), mRNAs encoding these proteins constitute frequent targets for DNA- and RNA-based gene therapy. Treatment of the DNA gels with BamHI results in the cleavage of the anti-HIV deozyribozyme from the gel, and downregulation of the Tat mRNA by the mechanism stated above. The HIV aptamer is synthesized in vitro using synthetic DNA templates under the control of promoters specific to T7 RNA polymerase. The aptamer is then radiolabeled using [5′-32P]pCp and T4 RNA ligase and purified on a 12% poly(acrylamide-co-bisacrylamide) non-denaturing gel. 3′-end labeled HIV aptamer is incubated along with the DNA gels and BamHI under physiological conditions, and the cleaved RNA fragments are analyzed by electrophoresis on a denaturing gel. Autoradiograms are quantified using Typhoon phosphoimager and ImageQuant software.
  • Physiological validation of the platform: Downregulation of miRNA for Age-Related Macular Degeneration. The physiological relevance of this platform in the context of developmental biology and clinical intervention are demonstrated by following similar steps to release an anti-miRNA ribozyme. The double stranded helix substrate includes a siRNA construct which targets and interferes with the functioning of the vascular endothelial growth factor receptor (VEGFR). Treatment of the DNA gels with BamHI results in the cleavage of the siRNA from the gel, and downregulation of the VEGFR mRNA. The VEGFR mRNA is synthesized in vitro using synthetic DNA templates under the control of promoters specific to T7 RNA polymerase. The aptamer is then radiolabeled using [5′-32P]pCp and T4 RNA ligase and purified on a 12% poly(acrylamide-co-bisacrylamide) non-denaturing gel. 3′-end labeled VEGFR aptamer is incubated along with the DNA gels and BamHI under physiological conditions, and the cleaved RNA fragments are analyzed by electrophoresis on a denaturing gel. Autoradiograms are quantified using Typhoon phosphoimager and ImageQuant software.
  • Kinetic Diffusion Studies in DNA Gels. Permeation studies are conducted using Valia-Chen diffusion cells consisting of donor and acceptor reservoirs with temperature control at 37° C. Each gel is pre-swollen in Tris buffer until equilibrium and placed between the two half diffusion cells. The transport of restriction endonuclease/cleaved fragments is determined via MALDI spectrophotomer. Permeability coefficients, partition coefficients, and diffusion coefficients are calculated. Comparison of the relative rates of diffusion between the influx of BamHI and the exodus of the cleaved fragments will yield information regarding the rate limiting steps of the reactive/diffusion process.
  • Structural Analysis. Equilibrium weight and volume swelling studies are conducted using conventional methods to calculate polymer fractional content, gel structural properties such as molecular weight between crosslinks, and correlation length (e.g., mesh size of the gel).
  • OBJECTIVE 2: The rational design, synthesis, characterization, and optimization of novel biohybrid hydrogel carriers which exhibit modulatory, on-off, release of a therapeutic via reversible aptamer-analyte crosslinking.
    • Specific Aims: 1) Perform in-vitro transcription and purification of a biotin-specific aptamer and characterize via equilibrium and kinetic binding isotherms; 2) Synthesize novel recognitive RNA-based hydrogels using aptamer and bioconjugate chemistry; 3) Elucidate the associated mesh size changes and swelling transitions via metal ion/chelating agent switching; 4) Perform dynamic release kinetics of a fluorescent molecule from the hydrogel carrier. Experiments:
  • In-vitro Transcription and Purification of an Analyte Specific Aptamer. Biotin-binding RNA pseudoknots are synthesized in vitro using synthetic DNA templates under the control of promoters specific to T7 RNA polymerase. Large quantities of DNA are generated using the Klenow fragment of DNA Polymerase I. Integrity of the transcripts are examined by denaturing polyacrylamide gel electrophoresis. Biotin binding pseudoknots are optimized for binding efficiency using SELEX. Pseudoknots are radiolabeled using [5′-32P]pCp and T4 RNA ligase and purified on a 12% poly(acrylamide-co-bisacrylamide) non-denaturing gel. Modified nucleotides are co-transcriptionally incorporated into RNA pseudoknots to render them resistant against ribonucleases and then selected for binding via affinity chromatography. For example, by modulating incorporation of fluorine-pyrimidine nucleotides (Epicentre biotechnologies) small RNAs resilient against degradation by ubiquitous RNAse A, can be produced. Protection from other RNases is achieved by posttranscriptional modification of accessible nucleotides with nucleotide specific reagents (kethoxal, DMS, DEPC).
  • Affinity Elution Studies. Binding properties of RNA aptamers are monitored using modified Sepharoses. Columns are equilibrated with Biotin Binding Buffer (20 mM HEPES, 100 mM KCl, 15 mM MgCl2). Loading the column with samples or magnesium/EDTA buffers is followed by spinning at 1000 rpm for 20 seconds.
  • Synthesis of Novel Recognitive RNA based Hydrogels using Aptamer and Bioconjugate Chemistry. Poly(acrylic acid-g-RNA-g-biotin) networks are prepared from poly(acrylic acid) chains of high monodispersity. Grafting of RNA and modified biotin amines to the matrix occur via commercially available cross-linking reagents (Maleimide chemistry-EDC, oxene chemistry). Hydrogels are assembled in polystyrene molds of precise dimensions.
  • Elucidation of Mesh Size Changes/Swelling Transitions via Metal Ion/Chelating Agent Switching. Observable macroscopic volume transitions are induced by metal ion/chelating agent switching, which are observed under an optical microscope. Gels are placed in known of volume of Tris buffer and the concentration of a FTIC Dextran (model drug) versus time at 37° C. will be used to determine modulatory release kinetics.
  • The present invention has been described in terms of specific embodiments incorporating details to facilitate the understanding of the principles of construction and operation of the invention. Such references herein to specific embodiments and details thereof are not intended to limit the scope of the claims appended hereto. It will be apparent to those skilled in the art that modifications can be made in the embodiments chosen for illustration without departing from the spirit and scope of the invention. Specifically, it will be apparent to one of ordinary skill in the art that the device of the present invention can be implemented in several different ways and the apparatus disclosed above is only illustrative of the preferred embodiment of the invention and is in no way a limitation.

Claims (23)

1. A method for making a drug delivery system, the method comprising:
a) providing backbone monomers, cross-linking monomers and nucleic acid moieties; and
b) initiating copolymerization of the backbone monomer and cross-linking monomer to thereby form a polymeric hydrogel with therapeutic units incorporated therein, wherein the therapeutic units include strands of the nucleic acid moieties.
2. The method of claim 1, further comprising forming a therapeutic dose from the polymeric hydrogel.
3. The method of claim 1, wherein forming a polymeric hydrogel comprises forming a solution comprising an amount of the strands of nucleic acid, the backbone monomer cross-linking monomer and initiating copolymerization of the backbone monomer and cross-linking monomer.
4. The method of claim 1, wherein the nucleic acid moieties include a drug.
5. The method of claim 4, wherein the drug is selected from the group consisting of an antibiotic, an anti-inflammatory, an antihistamine, an antiviral agent, a cancer drug, an anesthetic, a cycloplegic, a mydriatics, a lubricant agent, a hydrophilic agent, a decongestant, a vasoconstrictor, a vasodilater, an Immuno-suppressant, an immuno-modulating agent and an anti-glaucoma agent.
6. The method of claim 2, wherein the therapeutic dose is in a form of a pill or a patch.
7. A method of dispensing a therapeutic dose comprising:
a) administering a polymeric hydrogel with therapeutic units incorporated therein, wherein the therapeutic units include strands of nucleic acid; and
b) initiating a controlled release of portions of the therapeutic units.
8. The method of claim 7, wherein initiating the controlled release of the portions of the therapeutic units comprises one or more of heating the polymeric hydrogel, altering a pH polymeric hydrogel, altering an ionic strength of the polymeric hydrogel, exposing the polymeric hydrogel to an electric field and exposing the hydrogel to light.
9. The method of claim 7, wherein initiating the controlled release of the portions of the therapeutic units comprises treating the polymeric hydrogel with an enzyme.
10. The method of claim 7, wherein the polymeric hydrogel is placed in contact with a target tissue.
11. The method of claim 10, wherein the polymeric hydrogel is placed in contact with the target tissue by injecting the polymeric hydrogel into the target tissue.
12. The method of claim 7, wherein initiating the controlled release of the portions of the therapeutic units comprises treating the polymeric hydrogel with an enzyme.
13. The method of claim 7, wherein the therapeutic units comprise one or more active drugs.
14. The method of claim 13, wherein the one or more active drugs are selected from the group consisting of an antibiotic, an anti-inflammatory, an antihistamine, an antiviral agent, a cancer drug, an anesthetic, a cycloplegic, a mydriatics, a vasodilater, a lubricant agent, a hydrophilic agent, a decongestant, a vasoconstrictor, an immuno-suppressant, an immuno-modulating agent and an anti-glaucoma agent.
15. A system for delivering a therapeutic dose of an active drug, the system comprising a polymeric hydrogel with therapeutic units incorporated therein, wherein the therapeutic units include strands of the nucleic acid.
16. The system of claim 15, wherein therapeutic units include a drug moiety complex with the strands of nucleic acid.
17. The system of claim 16, wherein the drug is selected from the group consisting of an antibiotic, an anti-inflammatory, an antihistamine, an antiviral agent, a cancer drug, an anesthetic, a cycloplegic, a mydriatics, a lubricant agent, a hydrophilic agent, a decongestant, a vasoconstrictor, a vasodilater, an Immuno-suppressant, an immuno-modulating agent and an anti-glaucoma agent.
18. The system of claim 15, wherein the polymeric hydrogel is in a pill or a patch form.
19. The system of claim 15, wherein the therapeutic dose of a drug is controllably released from the polymeric hydrogel by enzymatic cleavage of the strands of the nucleic acid moieties.
20. A system for delivering a therapeutic dose of an active drug, the system comprising a delivery medium with therapeutic units incorporated therein, wherein the therapeutic units include strands of the nucleic acid.
21. The system of claim 20, wherein the delivery medium includes one or more of a gel matrix, metal particles, a polymer film, a polymer network, metal surface, polymer particles, particulate gels, particulate networks, a polymeric dendrimer and a surface conjugated with complexes.
22. The system of claim 20, wherein the delivery medium comprises metal nanoparticles with therapeutic nucleic acid strands and receptor moieties coupled thereto.
23. The system of claim 22, wherein the metal nanoparticles are silver or gold nanoparticles.
US11/985,263 2006-11-13 2007-11-13 Drug delivery system and method Abandoned US20080138408A1 (en)

Priority Applications (2)

Application Number Priority Date Filing Date Title
US11/985,263 US20080138408A1 (en) 2006-11-13 2007-11-13 Drug delivery system and method
US14/955,309 US10064953B2 (en) 2006-11-13 2015-12-01 Drug delivery system and method

Applications Claiming Priority (3)

Application Number Priority Date Filing Date Title
US85855306P 2006-11-13 2006-11-13
US93777307P 2007-06-28 2007-06-28
US11/985,263 US20080138408A1 (en) 2006-11-13 2007-11-13 Drug delivery system and method

Related Child Applications (1)

Application Number Title Priority Date Filing Date
US14/955,309 Continuation US10064953B2 (en) 2006-11-13 2015-12-01 Drug delivery system and method

Publications (1)

Publication Number Publication Date
US20080138408A1 true US20080138408A1 (en) 2008-06-12

Family

ID=39402250

Family Applications (2)

Application Number Title Priority Date Filing Date
US11/985,263 Abandoned US20080138408A1 (en) 2006-11-13 2007-11-13 Drug delivery system and method
US14/955,309 Expired - Fee Related US10064953B2 (en) 2006-11-13 2015-12-01 Drug delivery system and method

Family Applications After (1)

Application Number Title Priority Date Filing Date
US14/955,309 Expired - Fee Related US10064953B2 (en) 2006-11-13 2015-12-01 Drug delivery system and method

Country Status (2)

Country Link
US (2) US20080138408A1 (en)
WO (1) WO2008060575A2 (en)

Cited By (15)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US20100239637A1 (en) * 2008-12-11 2010-09-23 Massachusetts Institute Of Technology Contact lens drug delivery device
KR101255149B1 (en) 2011-10-14 2013-04-22 포항공과대학교 산학협력단 Composition for nucleic acid delivery using metal nanoparticles and preparing method thereof
US20130196915A1 (en) * 2010-01-23 2013-08-01 Yong Wang Affinity hydrogels for controlled protein release
US20140036225A1 (en) * 2012-07-31 2014-02-06 Khaled Chehab Lens incorporating myopia control optics and muscarinic agents
US9522241B2 (en) 2012-03-22 2016-12-20 Raytheon Company Controlled release nanoparticulate matter delivery system
US10106794B2 (en) * 2014-10-09 2018-10-23 The Research Foundation For The State University Of New York RNA hydrogel
US10206813B2 (en) 2009-05-18 2019-02-19 Dose Medical Corporation Implants with controlled drug delivery features and methods of using same
US10245178B1 (en) 2011-06-07 2019-04-02 Glaukos Corporation Anterior chamber drug-eluting ocular implant
US10406029B2 (en) 2001-04-07 2019-09-10 Glaukos Corporation Ocular system with anchoring implant and therapeutic agent
US10815486B2 (en) 2017-07-05 2020-10-27 The Research Foundation For The State University Of New York Chemically modified AMPA receptor RNA aptamers
US10959941B2 (en) 2014-05-29 2021-03-30 Glaukos Corporation Implants with controlled drug delivery features and methods of using same
US20220001024A1 (en) * 2019-11-01 2022-01-06 The Florida International University Board Of Trustees Guanylurea functionalized peptides and proteins for therapeutics
US11318043B2 (en) 2016-04-20 2022-05-03 Dose Medical Corporation Bioresorbable ocular drug delivery device
US11564833B2 (en) 2015-09-25 2023-01-31 Glaukos Corporation Punctal implants with controlled drug delivery features and methods of using same
US11925578B2 (en) 2015-09-02 2024-03-12 Glaukos Corporation Drug delivery implants with bi-directional delivery capacity

Families Citing this family (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US10285943B2 (en) 2010-12-02 2019-05-14 Greenmark Biomedical Inc. Aptamer bioconjugate drug delivery device
US10429387B2 (en) * 2016-07-27 2019-10-01 Universiteit Tweate Simple and affordable method for immuophenotyping using a microfluidic chip sample preparation with image cytometry
US10993910B2 (en) 2016-12-07 2021-05-04 Seoul National University R&Db Foundation Biopatch, bioheater, biosensor and bioelectronic patch device
JP2021519310A (en) 2018-03-28 2021-08-10 グリーンマーク バイオメディカル インコーポレイテッドＧｒｅｅｎＭａｒｋ Ｂｉｏｍｅｄｉｃａｌ， Ｉｎｃ． Phosphate cross-linked starch nanoparticles and dental treatment

Citations (25)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US4668506A (en) * 1985-08-16 1987-05-26 Bausch & Lomb Incorporated Sustained-release formulation containing and amino acid polymer
US4775531A (en) * 1985-05-29 1988-10-04 Eye Research Institute Of Retina Foundation Non-toxic opthalmic preparations
US4931279A (en) * 1985-08-16 1990-06-05 Bausch & Lomb Incorporated Sustained release formulation containing an ion-exchange resin
US5263888A (en) * 1992-02-20 1993-11-23 Matsushita Electric Industrial Co., Ltd. Method of manufacture of liquid crystal display panel
US5318780A (en) * 1991-10-30 1994-06-07 Mediventures Inc. Medical uses of in situ formed gels
US5568297A (en) * 1992-05-25 1996-10-22 Sharp Kabushiki Kaisha Method of making a liquid crystal display device by measuring the liquid crystal layer thickness and adjusting
US5711989A (en) * 1992-11-19 1998-01-27 Nordson Corporation Computer controlled method for dispensing viscous fluid
US5840338A (en) * 1994-07-18 1998-11-24 Roos; Eric J. Loading of biologically active solutes into polymer gels
US5876709A (en) * 1997-05-26 1999-03-02 New Vision Co., Ltd. Ophthalmic composition containing active Vitamin D
US6129804A (en) * 1998-05-01 2000-10-10 International Business Machines Corporation TFT panel alignment and attachment method and apparatus
US6222603B1 (en) * 1998-01-13 2001-04-24 Matsushita Electric Industrial Co., Ltd. Method of manufacturing liquid crystal display device with a double seal
US20010006968A1 (en) * 1999-12-23 2001-07-05 Julian Trimming Use of ophthalmic agent
US6329108B1 (en) * 1997-07-28 2001-12-11 Canon Kabushiki Kaisha Color-filter manufacturing method
US6375973B2 (en) * 2000-01-25 2002-04-23 Alcon Universal Ltd. Ophthalmic anti-allergy compositions suitable for use with contact lenses
US6541063B1 (en) * 1999-11-04 2003-04-01 Speedline Technologies, Inc. Calibration of a dispensing system
US6552103B1 (en) * 1997-09-03 2003-04-22 The Regents Of The University Of California Biomimetic hydrogel materials
US6703039B2 (en) * 2000-12-06 2004-03-09 Bausch & Lomb Incorporated Reversible gelling system for ocular drug delivery
US6730065B1 (en) * 2000-09-15 2004-05-04 Ocularis Pharma, Inc. Night vision composition
US6735470B2 (en) * 2000-05-31 2004-05-11 Biophoretic Therapeutic Systems, Llc Electrokinetic delivery of medicaments
US20040096477A1 (en) * 2002-06-05 2004-05-20 Anuj Chauhan Ophthalmic drug delivery system
US20050163844A1 (en) * 2004-01-26 2005-07-28 Control Delivery Systems, Inc. Controlled and sustained delivery of nucleic acid-based therapeutic agents
US20050208102A1 (en) * 2003-04-09 2005-09-22 Schultz Clyde L Hydrogels used to deliver medicaments to the eye for the treatment of posterior segment diseases
US20060100408A1 (en) * 2002-03-11 2006-05-11 Powell P M Method for forming contact lenses comprising therapeutic agents
US20060177483A1 (en) * 2005-02-04 2006-08-10 Byrne Mark E Contact drug delivery system
US7332586B2 (en) * 2001-07-10 2008-02-19 North Carolina State University Nanoparticle delivery vehicle

Patent Citations (29)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US4775531A (en) * 1985-05-29 1988-10-04 Eye Research Institute Of Retina Foundation Non-toxic opthalmic preparations
US4911933A (en) * 1985-05-29 1990-03-27 Eye Research Institute Of Retina Foundation Non-toxic opthalmic preparations
US4931279A (en) * 1985-08-16 1990-06-05 Bausch & Lomb Incorporated Sustained release formulation containing an ion-exchange resin
US4668506A (en) * 1985-08-16 1987-05-26 Bausch & Lomb Incorporated Sustained-release formulation containing and amino acid polymer
US5958443A (en) * 1991-10-30 1999-09-28 Mdv Technologies, Inc. Medical uses of in situ formed gels
US5318780A (en) * 1991-10-30 1994-06-07 Mediventures Inc. Medical uses of in situ formed gels
US5587175A (en) * 1991-10-30 1996-12-24 Mdv Technologies, Inc. Medical uses of in situ formed gels
US6136334A (en) * 1991-10-30 2000-10-24 Mdv Technologies, Inc. Medical uses of in situ formed gels
US5263888A (en) * 1992-02-20 1993-11-23 Matsushita Electric Industrial Co., Ltd. Method of manufacture of liquid crystal display panel
US5568297A (en) * 1992-05-25 1996-10-22 Sharp Kabushiki Kaisha Method of making a liquid crystal display device by measuring the liquid crystal layer thickness and adjusting
US5711989A (en) * 1992-11-19 1998-01-27 Nordson Corporation Computer controlled method for dispensing viscous fluid
US5840338A (en) * 1994-07-18 1998-11-24 Roos; Eric J. Loading of biologically active solutes into polymer gels
US5876709A (en) * 1997-05-26 1999-03-02 New Vision Co., Ltd. Ophthalmic composition containing active Vitamin D
US6329108B1 (en) * 1997-07-28 2001-12-11 Canon Kabushiki Kaisha Color-filter manufacturing method
US6552103B1 (en) * 1997-09-03 2003-04-22 The Regents Of The University Of California Biomimetic hydrogel materials
US6222603B1 (en) * 1998-01-13 2001-04-24 Matsushita Electric Industrial Co., Ltd. Method of manufacturing liquid crystal display device with a double seal
US6129804A (en) * 1998-05-01 2000-10-10 International Business Machines Corporation TFT panel alignment and attachment method and apparatus
US6541063B1 (en) * 1999-11-04 2003-04-01 Speedline Technologies, Inc. Calibration of a dispensing system
US20010006968A1 (en) * 1999-12-23 2001-07-05 Julian Trimming Use of ophthalmic agent
US6375973B2 (en) * 2000-01-25 2002-04-23 Alcon Universal Ltd. Ophthalmic anti-allergy compositions suitable for use with contact lenses
US6735470B2 (en) * 2000-05-31 2004-05-11 Biophoretic Therapeutic Systems, Llc Electrokinetic delivery of medicaments
US6730065B1 (en) * 2000-09-15 2004-05-04 Ocularis Pharma, Inc. Night vision composition
US6703039B2 (en) * 2000-12-06 2004-03-09 Bausch & Lomb Incorporated Reversible gelling system for ocular drug delivery
US7332586B2 (en) * 2001-07-10 2008-02-19 North Carolina State University Nanoparticle delivery vehicle
US20060100408A1 (en) * 2002-03-11 2006-05-11 Powell P M Method for forming contact lenses comprising therapeutic agents
US20040096477A1 (en) * 2002-06-05 2004-05-20 Anuj Chauhan Ophthalmic drug delivery system
US20050208102A1 (en) * 2003-04-09 2005-09-22 Schultz Clyde L Hydrogels used to deliver medicaments to the eye for the treatment of posterior segment diseases
US20050163844A1 (en) * 2004-01-26 2005-07-28 Control Delivery Systems, Inc. Controlled and sustained delivery of nucleic acid-based therapeutic agents
US20060177483A1 (en) * 2005-02-04 2006-08-10 Byrne Mark E Contact drug delivery system

Non-Patent Citations (4)

* Cited by examiner, † Cited by third party
Title
Elghanian, et al. (1997) "Selective Colorimetric Detection of Polynucleotides Based on the Distance-Dependent Optical Properties of Gold Nanoparticles", Science, 277: 1078-81. *
Eric Berger, http://blog.chron.com/sciguy/2010/10/in-all-the-world-how-many-diseases-have-humans-succeeded-in-eradicating/, Huston Chronicle, Huston TX, posted online October 20, 2010, downloaded 5/21/15, 1 page printout. *
Kulbanchinskiy (2007) "Methods for Selection of Aptamers to Protein Targets", Biochemistry (Moscow), 72(13): 1505-18. *
Taton, et al. (2000) "Scanometric DNA Array Detection With Nanoparticle Probes", Science, 289: 1757-60. *

Cited By (21)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US10406029B2 (en) 2001-04-07 2019-09-10 Glaukos Corporation Ocular system with anchoring implant and therapeutic agent
US8414912B2 (en) * 2008-12-11 2013-04-09 Massachusetts Institute Of Technology Contact lens drug delivery device
US20100239637A1 (en) * 2008-12-11 2010-09-23 Massachusetts Institute Of Technology Contact lens drug delivery device
US10206813B2 (en) 2009-05-18 2019-02-19 Dose Medical Corporation Implants with controlled drug delivery features and methods of using same
US11426306B2 (en) 2009-05-18 2022-08-30 Dose Medical Corporation Implants with controlled drug delivery features and methods of using same
US20130196915A1 (en) * 2010-01-23 2013-08-01 Yong Wang Affinity hydrogels for controlled protein release
US10245178B1 (en) 2011-06-07 2019-04-02 Glaukos Corporation Anterior chamber drug-eluting ocular implant
US8747903B2 (en) 2011-10-14 2014-06-10 Postech Academy-Industry Foundation Composition for nucleic acid delivery using metal nanoparticles and preparing method thereof
KR101255149B1 (en) 2011-10-14 2013-04-22 포항공과대학교 산학협력단 Composition for nucleic acid delivery using metal nanoparticles and preparing method thereof
US9522241B2 (en) 2012-03-22 2016-12-20 Raytheon Company Controlled release nanoparticulate matter delivery system
US10569067B2 (en) * 2012-03-22 2020-02-25 Raytheon Company Controlled release nanoparticulate matter delivery system
US9827250B2 (en) * 2012-07-31 2017-11-28 Johnson & Johnson Vision Care, Inc. Lens incorporating myopia control optics and muscarinic agents
US20140036225A1 (en) * 2012-07-31 2014-02-06 Khaled Chehab Lens incorporating myopia control optics and muscarinic agents
US11253394B2 (en) 2013-03-15 2022-02-22 Dose Medical Corporation Controlled drug delivery ocular implants and methods of using same
US10959941B2 (en) 2014-05-29 2021-03-30 Glaukos Corporation Implants with controlled drug delivery features and methods of using same
US10106794B2 (en) * 2014-10-09 2018-10-23 The Research Foundation For The State University Of New York RNA hydrogel
US11925578B2 (en) 2015-09-02 2024-03-12 Glaukos Corporation Drug delivery implants with bi-directional delivery capacity
US11564833B2 (en) 2015-09-25 2023-01-31 Glaukos Corporation Punctal implants with controlled drug delivery features and methods of using same
US11318043B2 (en) 2016-04-20 2022-05-03 Dose Medical Corporation Bioresorbable ocular drug delivery device
US10815486B2 (en) 2017-07-05 2020-10-27 The Research Foundation For The State University Of New York Chemically modified AMPA receptor RNA aptamers
US20220001024A1 (en) * 2019-11-01 2022-01-06 The Florida International University Board Of Trustees Guanylurea functionalized peptides and proteins for therapeutics

Also Published As

Publication number Publication date
US20160082124A1 (en) 2016-03-24
WO2008060575A2 (en) 2008-05-22
WO2008060575A3 (en) 2008-11-13
US10064953B2 (en) 2018-09-04

Similar Documents

Publication Publication Date Title
US10064953B2 (en) Drug delivery system and method
Mo et al. DNA hydrogel-based gene editing and drug delivery systems
Hu et al. Hydrogel contact lens for extended delivery of ophthalmic drugs
Singh et al. Hydrogels: a review
Pillai et al. Polymers in drug delivery
EP1848749B1 (en) Contact drug delivery system
Zelikin et al. A general approach for DNA encapsulation in degradable polymer microcapsules
Coelho et al. Drug delivery systems: Advanced technologies potentially applicable in personalized treatments
Tashakori-Sabzevar et al. Development of ocular drug delivery systems using molecularly imprinted soft contact lenses
Truong et al. Self-catalyzed degradable cationic polymer for release of DNA
WO2003022910A1 (en) Synthesis and uses of polymer gel nanoparticle networks
MX2010012236A (en) Polymeric carrier.
JP5969164B2 (en) Polymer-siRNA nanoparticle carrier simultaneously linked by charge bond and biodegradable covalent bond and method for producing the same
Flessner et al. Degradable polyelectrolyte multilayers that promote the release of siRNA
Jia et al. Self-assembly of DNA-containing copolymers
US8349352B2 (en) Therapeutic contact lenses with anti-fungal delivery
Lin et al. Grafting from small interfering ribonucleic acid (siRNA) as an alternative synthesis route to siRNA–polymer conjugates
WO2009032266A2 (en) Dispersions of microparticles and microgels in hydrogels for drug delivery
Chevalier si RNA Targeting and Treatment of Gastrointestinal Diseases
Datta Characterization of polyethylene glycol hydrogels for biomedical applications
WO2012144560A1 (en) Self-gelatinizable nucleic acid
Jeong et al. RNA-polymer hybrids via direct and site-selective acylation with the ATRP initiator and photoinduced polymerization
Li et al. Rational design of dna hydrogels based on molecular dynamics of polymers
Baldwin et al. DNA-crosslinked alginate and layered microspheres to modulate the release of encapsulated FITC-dextran
Venkatesh et al. Nucleic acid therapeutic carriers with on-demand triggered release

Legal Events

Date Code Title Description
AS Assignment

Owner name: AUBURN UNIVERSITY, ALABAMA

Free format text: ASSIGNMENT OF ASSIGNORS INTEREST;ASSIGNORS:VENKATESH, SIDDARTH;WOWER, JACEK;BYRNE, MARK E.;REEL/FRAME:020528/0508

Effective date: 20080117

STCB Information on status: application discontinuation

Free format text: ABANDONED -- FAILURE TO RESPOND TO AN OFFICE ACTION