EP1206285A2 - Complexes vecteurs de medicament et technique d'utilisation de ceux-ci - Google Patents

Complexes vecteurs de medicament et technique d'utilisation de ceux-ci

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Publication number
EP1206285A2
EP1206285A2 EP00955415A EP00955415A EP1206285A2 EP 1206285 A2 EP1206285 A2 EP 1206285A2 EP 00955415 A EP00955415 A EP 00955415A EP 00955415 A EP00955415 A EP 00955415A EP 1206285 A2 EP1206285 A2 EP 1206285A2
Authority
EP
European Patent Office
Prior art keywords
nucleotide
drug
carrier
drag
component
Prior art date
Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
Withdrawn
Application number
EP00955415A
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German (de)
English (en)
Inventor
Mikhail I. Papisov
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.)
General Hospital Corp
Original Assignee
General Hospital Corp
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Filing date
Publication date
Application filed by General Hospital Corp filed Critical General Hospital Corp
Publication of EP1206285A2 publication Critical patent/EP1206285A2/fr
Withdrawn legal-status Critical Current

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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
    • A61K47/51—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
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P31/00—Antiinfectives, i.e. antibiotics, antiseptics, chemotherapeutics
    • A61P31/04—Antibacterial agents
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P31/00—Antiinfectives, i.e. antibiotics, antiseptics, chemotherapeutics
    • A61P31/12—Antivirals
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P33/00—Antiparasitic agents
    • A61P33/02—Antiprotozoals, e.g. for leishmaniasis, trichomoniasis, toxoplasmosis
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P35/00—Antineoplastic agents
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P43/00—Drugs for specific purposes, not provided for in groups A61P1/00-A61P41/00

Definitions

  • nucleic acid sequences in cells e.g., DNA of a cancerous cell
  • drugs employed to treat diseases are either insufficiently soluble in aqueous solutions or have adverse side effects, such as the death of healthy cells, because of the lack of suitable substances to deliver drugs to a cell or organism (e.g., mammal) requiring treatment.
  • macromolecular drug-carriers which most commonly are water-soluble macromolecules with chemically associated drug molecules, often are employed to prolong drug circulation, limit renal clearance, increase drug accumulation in target tissues or cells, and to decrease drug concentration in normal tissues.
  • Several model and prototype carriers of this type have been developed. Potentially, these carriers can be as small as 5-10 nanometers (nm), but depending on the drug structure and content, they often form larger (20-50 nm) associates. Carriers of this type are intended to act, essentially, as pro-drugs, the drug substance as a result of degradation of the drug-carrier bond. Some carriers of this type have been targeted to cancer cell markers.
  • Examples of this class of drug-carriers are: dextran-mitomycin conjugates; HPMA-doxorubicin conjugates with enzyme-degradable peptide bonds between the drug molecule and the backbone polymer; doxorubicin-Fab conjugates with pH-sensitive bonds between doxorubicin molecules and the Fab.
  • carriers of this type have at least two potential drawbacks.
  • drug release via degradation of the drug-carrier bond generally is irreversible.
  • drug released from the carrier will circulate in the body independently of the carrier, which may reduce the efficacy of drug delivery.
  • Drug release via enzyme-dependent or pH-dependent hydrolysis has been reported to improve the ratio of drug activity in the target relative to normal tissues.
  • expression of enzymes, such as proteases in tumors and other pathologies is highly variable, which makes predictability of release rate of the drug difficult.
  • Enzyme- independent biodegradation can occur in both pathological and normal tissues.
  • a second problem relates to exposure to the environment of drug molecules attached to the macromolecular backbone. This can result in cross-interaction of drug moieties with formation of intramolecular and intermolecular micelles, interactions with tissue components altering drug-carrier adduct biodistribution, and other undesirable effects. These effects are expected to be partially suppressed via "steric protection,” or modification of the carrier backbone with hydrophilic polymer chains such as, for example, polyethyleneglycol, dextran, or PHF
  • Microparticles and emulsions were developed as an alternative where the drug molecules are not bound chemically, but rather are adsorbed on, or dissolved in, the material of the carrier.
  • particles and emulsions do not circulate in vivo long enough and accumulate in the reticuloendothelial system (RES) and other organs, unless the particle (droplet) surface is modified with a hydrophilic polymer, such as PEG.
  • RES reticuloendothelial system
  • PEG reticuloendothelial system
  • the overall size of sterically protected particles (droplets) is usually above about 25 nm.
  • Major problems in the development of such carriers include the fact that (1 ) the emulsions generally are relatively unstable and change (e.g.
  • micelles which were developed as "self-assembling" drug carriers similar to particles and emulsions. They are made of surfactants, which are usually block copolymers, where one of the blocks is hydrophilic, and the other hydrophobic. The total hydrodynamic size of the micelles usually is 10-30 nm.
  • the hydrophobic drug molecules either are incorporated into the hydrophobic core or, alternatively, chemically conjugated with one of the blocks and form the hydrophobic core.
  • Still another attempt includes encapsulation of drugs in the aqueous compartments of liposomes, which are vesicles, typically having a diameter in a range of between about 50 and about 1000 nm.
  • liposomes which are vesicles, typically having a diameter in a range of between about 50 and about 1000 nm.
  • drug encapsulation and the potential to control drug delivery by incorporation into liposomes can be problematic.
  • drug release from liposomes generally is irreversible.
  • liposome penetration into tumors or tumor zones that have relatively low vascular permeability often is poor.
  • there are many problems associated with high- volume production and storage of liposomal preparation that present significant technical challenges.
  • the present invention relates to the field of drug delivery, in particular to methods of forming drug-carrier complexes and the use of drug-carrier complexes as pharmaceutical compositions to deliver and target drugs in an organism, tissue culture or cells.
  • the method includes forming a drug-carrier complex by combining at least one nucleotide strand with a drug, whereby the drug and the nucleotide strand reversibly associate with each other to form a drug-carrier complex.
  • the method includes forming a drug-carrier complex by combining a drug with at least two nucleotide strands that hybridize with each other, whereby the drug associates with the nucleotide strands to form a water soluble drug-carrier complex.
  • the method includes forming a drug-carrier composition by combining a drug component and a nucleotide component.
  • the combined drug and nucleotide components are lyophilized to form the drug-carrier composition.
  • the method includes forming a drug-carrier composition by lyophilizing a drug component, lyophilizing a nucleotide component and combining the lyophilized drug component and the lyophilized nucleotide component to form the drug-carrier composition.
  • Another embodiment of the invention is a drug carrier, comprising a double- stranded nucleotide and a polymer component covalently bonded to at least one strand of the double-stranded nucleotide.
  • the polymer component has an aqueous solubility of at least one mg/liter at 25°C.
  • An additional embodiment of the invention is a drug carrier, comprising a double-stranded nucleotide and an oligomer component covalently bonded to at least one strand of the double-stranded nucleotide.
  • the invention is a drug-carrier complex, comprising a single-stranded nucleotide, a drug reversibly associated with the single-stranded nucleotide and a polymer associated with the drug or the single- stranded nucleotide.
  • the invention is a drug-carrier complex, comprising a single-stranded nucleotide, an oligomer associated with the single- stranded nucleotide and a drug reversibly associated with the oligomer or the single- stranded nucleotide.
  • the invention is a drug carrier, comprising a single-stranded nucleotide and at least two polymers associated with the single- stranded nucleotide.
  • the invention is a drug carrier, comprising an oligomer, a single-stranded nucleotide entrapped by the oligomer and a drug reversibly associated with the single-stranded nucleotide.
  • the invention is a drug-carrier composition, comprising a nucleotide carrier component and a drug component.
  • the drug-carrier composition has a moisture content less than about 5% by weight.
  • the invention includes a drug-carrier composition consisting essentially of a drug component and a nucleotide component.
  • the invention is a pharmaceutical formulation, comprising a nucleotide carrier component and a drug in reversible association with the nucleotide carrier component.
  • a method of the invention includes delivering a drug to an organism by administering a drug-carrier complex to the organism.
  • the drug-carrier complex includes a nucleotide carrier and a drug in reversible association with each other.
  • a method of the invention includes delivering a drug to a tissue culture by administering a drug-carrier complex to the tissue culture.
  • the drug- carrier complex includes a nucleotide carrier and a drug in reversible association with each other.
  • the method includes delivering a drug to an organism by administering a drug and a nucleotide carrier, which reversibly associates with the drug to form a drug-carrier complex, to the organism.
  • the method includes delivering a drug to an organism by forming a drug carrier complex that includes a drug and a nucleotide strand in reversible association with the drug and administering the drug-carrier complex to the organism.
  • Another embodiment includes a method of delivering a drug to an organism by administering to the organism a drug-carrier complex.
  • the drug-carrier complex includes a drug component and a carrier component in reversible association with each other.
  • the drug can dissociate from the drug-carrier complex and reassociate with the carrier component.
  • the degree of association can depend, for example, on the concentrations of the drug and the carrier.
  • the method includes increasing aqueous solubility of a substance by reversibly associating the substance with a nucleotide carrier to form a water-soluble complex.
  • the invention is a targeted carrier, comprising a nucleotide, a polymer component associated with the nucleotide, and a ligand associated with the nucleotide or the polymer component and associable with a cell or tissue marker.
  • the cell or tissue marker is selected from the group consisting of proteins, peptides, carbohydrates, lipids and nucleotides.
  • the invention relates to a targeted carrier, comprising a nucleotide, a polymer component associated with the nucleotide and a ligand.
  • the ligand is associated with the nucleotide or the polymer component and is associable with a cell or tissue marker.
  • the cell or tissue marker is selected from the group consisting of proteins, peptides, carbohydrates, lipids and nucleotides.
  • the invention relates to a targeted drug-carrier complex, comprising a nucleotide, a drug reversibly associated with the nucleotide and a targeting component.
  • the targeting component is associated with the nucleotide or the drug.
  • the targeting component includes a ligand associable with a cell or tissue marker.
  • the cell or tissue marker is selected from the group consisting of proteins, peptides, carbohydrates, lipids and nucleotides.
  • the invention relates to a targeted drug-carrier complex, comprising a nucleotide, a drug reversibly associated with the nucleotide, a polymer component associated with the nucleotide or the drug and a targeting component.
  • the targeting component is associated with the nucleotide, the drug or the polymer.
  • the targeting component includes a ligand associable with a cell or tissue marker.
  • the cell or tissue marker is selected from the group consisting of proteins, peptides, carbohydrates, lipids and nucleotides.
  • the invention relates to a drug delivery system, comprising a matrix, a nucleotide associated with or entrapped within the matrix and a drug in reversible association with the nucleotide.
  • Another embodiment includes an implant, comprising an implant matrix, a nucleotide associated with or entrapped within the matrix and a drug in reversible association with the nucleotide.
  • the invention described herein provides drug-carrier complexes, drug-carrier compositions, drug carriers, pharmaceutical formulations, methods of delivery drugs to organisms and tissue cultures, targeted carriers and implants to deliver drugs to an organism, a tissue culture or a combination of cells.
  • the nucleotide-based drug delivery systems of the present invention have many advantages. For example, they can transport drugs in chemically unmodified form, and can reabsorb the released drug. By employing a reversible drug association, the drug delivery systems of this invention are able to reincorporate the released drug.
  • drug behavior in the tissues may remain dependent on the drug release system for as long as the latter remains functional, which offers the possibility of new opportunities in regulation of pharmacokinetics and pharmacodynamics. In a clinical setting, this is expected to result in better biological functionality and broader safety margins of pharmaceutical formulations and devices.
  • a relatively small size of the drug- carrier complex such as, for example, about 3 or about 5 nm.
  • the drug-carrier complex can be, for example, 5 to 10 times smaller than polymer- and micelle-based carriers, and at least 10 to 20 times smaller than liposomes. Therefore, drug penetration into certain tissues, such as cancerous tissues, may be significantly more efficient, especially where endothelial and interstitial barriers are high. Also, stability and release rates of drug-carrier complexes of the invention can be controlled within a broad range, thereby providing the opportunity to design products in accordance with specific clinical objectives.
  • release of drugs by the drug-carrier complexes of the invention does not require interactions with enzymes, cells or other factors, thereby making the drug-carrier complexes more independent of the organism and tissue state.
  • complexes of the invention can be designed to exploit specific conditions of an organism or tissue state, such as pH or enzyme content.
  • the components of the drug-carrier complexes of the invention can be made of close analogs of natural components of biological systems which are known to be completely biodegradable and non-toxic. Other specific advantages of the invention include the possibility of steric protection against carrier clearance and drug inactivation.
  • the drug-carrier complexes of the invention generally have no problems relevant to intramolecular or intermolecular association of drug molecules.
  • methods of forming and processing the drug-carrier complexes of the invention are readily scalable.
  • drug-carrier complexes are lyophilizable, and all components of the complexes can be stable in the presence of air.
  • no toxic surfactants are employed, the size of the complexes generally is stable and does not depend on conditions and concentration.
  • Drug release rate within an organism generally does not depend on highly variable adsorption forces. Ultrafiltration typically does not affect size and structure of the drug-carrier complexes.
  • Figure 1 A depicts the intercalation of doxorubicin (darkly shaded) with B- helical DNA in a space- filled model of one embodiment of the drug-carrier complex of the invention.
  • Figure IB depicts the intercalation of doxorubicin (darkly shaded) with B- helical DNA in a ball and stick model of another embodiment of the drug-carrier complex of the invention.
  • Figure 2 is a diagrammatic representation of a sterically protected embodiment of a drug-carrier complex of the invention.
  • Figures 3A, 3B, 3C, 3D and 3E depict additional embodiments of drug delivery complexes of the invention, portraying various arrangements of drug, polynucleotide or oligonucleotide, and polymer components of the complexes.
  • Figure 4 shows a schematic representation of a method of forming a drug delivery system of the invention by combining a drug with a gel matrix crosslinked through nucleotide strands that hybridize with each other.
  • the present invention relates to the discovery that useful drug-carrier complexes can be formed by combining a nucleotide (e.g., nucleotide strands) with a drug so that the drug and nucleotide are in reversible association with either other.
  • the drug-carrier complex can be used to deliver drugs to, for example, an organism, a tissue culture, or individual cells.
  • the invention further relates to the discovery that the drug-carrier complexes of the invention can be used in pharmaceutical formulations to increase the solubility of drugs, as targeted carriers, as drug delivery systems and as implants.
  • a drug-carrier complex is formed by combining at least one nucleotide strand with a drug.
  • the drug is in reversible association with the nucleotide component to form the drug-carrier complex.
  • a drug-carrier complex refers to at least one nucleotide strand and a drug that are in reversible association with each other.
  • association can be reversible, irreversible or both.
  • the association can be a physical association, a chemical association or both.
  • an association can be a covalent bond, a hydrophobic interaction, etc.
  • a "reversible association,” as defined herein, is an association wherein the components can return to an original, pre-association, state.
  • a reversible association of the components of a drug-carrier complex of the invention can disassociate and thereby return to original and distinct drug and nucleotide components.
  • the amount of association of components of a reversible association depends, at least in part, on the concentration of the drug and the carrier.
  • the components are dissociable under physiological conditions.
  • the reversible associations are associations selected from the group consisting of electrostatic bonding, hydrogen bonding, van der Waals forces, ionic interaction or donor/acceptor bonding.
  • the reversible association can be mediated by one or more associations between the drug and the nucleotide strand.
  • the reversible association can include a combination of hydrogen bonding and ionic bonding between the drug and the nucleotide strand.
  • the reversible association can be in combination with, for example, covalent or other noncovalent interactions between components, such as between a drug and a nucleotide.
  • a substance comprising a metal containing substance (e.g., platinum, cis-platinum, carboplatin, platinum, gold, silver) is reversibly associated with nucleotide carriers.
  • the association is considered to be non-covalent and can reversibly release a metal- containing biologically active drug component that can differ from the substance originally employed to form the drug-camer complex.
  • nucleotide As employed herein, the terms “nucleotide,” “nucleotide strand,” or “nucleotide carrier” describe a molecule consisting essentially of either naturally occurring nucleosides, (e.g., containing base components guanine (G), thymine (T), uracil (U), cytosine (C), adenine (A)), or their derivatives, or structural analogs.
  • a nucleotide strand comprises two or more nucleotides, e.g., an oligonucleotide, a polynucleotide, or a chemical derivative or an analog thereof.
  • oligonucleotide generally describes a molecule with well-defined structure and length (e.g., 5'-ACTTGCCATT, SEQ ID NO: 13).
  • polynucleotide generally refers to molecules assembled from a large number of nucleosides, where either the sequence or the length of the polynucleotide varies (e.g., preparations of DNA or RNA obtained from cell lysates, random polymers of the structure A m G k C,).
  • nucleotide strands may exist in linear and circular forms and are known to form a variety of structures, e.g., helical double strands (helixes), triple strands (often refened to as triplexes), loops, folds, crosses or supercoils.
  • This invention utilizes all types of nucleotide strands, structures and combinations formed thereof, including, for example, linear deoxyribonucleotides, linear ribonucleotides, linear ohgonucleotides comprising both ribonucleotides and deoxyribonucleotides, circular DNA (e.g, plasmids), folded ribonucleotides (e.g., ribozymes, t-RNA), viral RNA, viral DNA; DNA and RNA from cell lysates; synthetic polydeoxyribonucleotides and polyribonucleotides, chemically crosslinked double-stranded ohgonucleotides, partially or completely methylated or otherwise chemically altered forms of any of the above.
  • linear deoxyribonucleotides linear ribonucleotides, linear ohgonucleotides comprising both ribonucleotides and deoxyribonucleotides
  • circular DNA e
  • the prefened nucleotides of this invention are nucleotides with well-defined structures, such as synthetic ohgonucleotides, plasmids, RNA transcripts, viral DNA, and viral RNA (e.g., viral nucleotides in the viral envelope, intact virions, viruses).
  • nucleotides are synthetic ohgonucleotides chemically modified such that to modulate their biodegradation rate (e.g., ohgonucleotides comprising phosphorothioate linkages) or to enable conjugation with other molecules (for example, synthetic ohgonucleotides with a carboxyl or an amino group incorporated at 3'-end, 5'-end, or at one or more of the bases).
  • the DNA of a nucleotide strand can be B DNA (Drew, H.R., et al, Proc.
  • the nucleotides or nucleotide strands can be naturally occurring (e.g., isolated from cells of an organism, from tissue culture cells, a virus) or can be synthetically generated by, for example, a nucleotide synthesis apparatus.
  • the residues can be modified further after synthesizing the nucleotide strand.
  • the 5'-amino group of a nucleotide strand can be modified with N-hydroxy succinimide ester of carboxy-polyethyleneglycol.
  • Nucleosides typically occurring in nature can be used in conjunction with nucleosides not typically occurring in nature to synthesize the nucleotides and nucleotide strands employed by the invention.
  • nucleotide component refers to the drug-binding (drug carrying) component of the of the drug-carrier complexes of this invention.
  • the drug-binding component comprises at least one nucleotide strand.
  • the nucleotide component may include, or be further associated with, other components (e.g. polymers, oligomers, ligands) to form drug carrier, a drug delivery system, or a drug-laden implant.
  • the nucleotide strand is an oligonucleotide strand.
  • the drug of the drug-carrier complex can be any substance that binds reversibly (also refened to herein as "reversibly associates” or “is in reversible association") with a nucleotide or nucleotide strand, or any structures formed by said strands, of the invention.
  • the drug can reversibly associate with a single nucleotide of one or more nucleotide strands, for example via a donor-acceptor bond.
  • the drug can also reversibly associate with more than one nucleotide of one or more nucleotide strands.
  • the drug can reversibly associate with one nucleotide strand of a drug-carrier complex consisting of two nucleotide strands.
  • the drug can reversibly associate with two nucleotide strands of a drug-carrier complex consisting of two nucleotide strands.
  • the drug can reversibly associate with a single nucleotide strand of a drug-carrier complex consisting of three nucleotide strands.
  • the drug of the pharmaceutical formulation is a therapeutic drug.
  • therapeutic when referring to a drug used in the invention, refers to a drug used to treat, remediate or cure a disorder or a disease (e.g., hereditary diseases, viral diseases such as AIDS, cancer).
  • the drug of the pharmaceutical formulation is a diagnostic drug (e.g., a radioactive diagnostic drug, a flourescent diagnostic drug, a paramagnetic diagnostic drug, superparamagnetic diagnostic drug, an x-ray dense diagnostic drug or an electron dense diagnostic drug).
  • diagnosis when referring to a drug employed in the invention, refers to a drug employed to determine the nature or extent of a disease, or employed to confirm the presence of a disorder or a disease.
  • the drug can be, for example, an anticancer drug, antiviral drug, antibacterial drug, or antiprotozoal drug.
  • the drug can also be, for example, anthracycline, actinomycin, anthracenedione, bleomycin, mithramycin, chromomycin, olivomycin, protein, peptide, carbohydrate, polyamine, polycation, actinomycin D, daunorubicin, doxorubicin, idarubicin, bis-anthracycline, mitoxantrone, bleomycin A2, distamycin, netropsin, cisplatin, carboplatin, a silver ion and particle, or a gold ion and particle.
  • the drug is an oligonucleotide drug.
  • oligonucleotide drug refers to a molecule containing at least two nucleotides which binds reversibly to the nucleotide strand of the drug-carrier complex.
  • the oligonucleotide drug can be, for example, an antisense oligonucleotide or a ribozyme.
  • suitable drugs include a component such as a metal containing substance (e.g., platinum, cis-platinum, carboplatin, platinum, gold, silver), or a drug that binds the minor or major groove of DNA or RNA helix.
  • the drug includes at least one amino group.
  • the drug doxorubicin includes an amino group.
  • drug complex 16 includes a double-stranded oligonucleotide core 18 that carries a drug (not shown).
  • Polymers 20 are associated, such as by covalent bonding, with oligonucleotide core 18, thereby providing steric protection.
  • Figures 3 A through 3E represent additional embodiments of the drug-carrier complexes of the invention. Specifically, Figure 3 A shows micelle 22, that includes drug 24 reversibly associated with double-stranded ohgonucleotides 26. Polymers 28 are arcayed radially from double-stranded ohgonucleotides 26.
  • Figure 3B shows polymer-modified DNA 30, wherein drug 32 is reversibly associated with single- or double-stranded polynucleotide or oligonucleotide 34.
  • Polymers 36 extend from polynucleotide or oligonucleotide 34.
  • Figure 3C shows drug-carrier complex 38, wherein polymer backbone 40 is bound to multiple ohgonucleotides 42.
  • Drug component 44 is reversibly associated with ohgonucleotides 42, and polymers 46 extend from the backbone and sterically protect ohgonucleotides 42.
  • Figure 3D shows the drug-carrier complex as modified plasmid 48.
  • Drug 50 and polymer 52 are associated with each other, and the drug is reversibly associated with plasmid component 54.
  • Figure 3E shows the drug-carrier complex as gel particle 56, wherein ohgonucleotides 58 and associated drug molecules 60 are entrapped in gel 62.
  • Polymers 64 extend from gel 62.
  • the drug-carrier complexes of the invention include at least one drug.
  • a drug-carrier complex can include at least one nucleotide strand reversibly associated with an oligonucleotide drug (e.g., a ribozyme, an antisense oligonucleotide), an antibacterial drug and a metal containing substance (e.g., platinum, cis-platinum, carboplatin, platinum, gold, silver).
  • an oligonucleotide drug e.g., a ribozyme, an antisense oligonucleotide
  • an antibacterial drug e.g., an antibacterial drug and a metal containing substance (e.g., platinum, cis-platinum, carboplatin, platinum, gold, silver).
  • a metal containing substance e.g., platinum, cis-platinum, carboplatin, platinum, gold, silver.
  • the drug of the drug-carrier complex is combined with at least two nucleotide strands which hybridize with each other in the drug-carrier complexes of the invention.
  • a second nucleotide strand is combined with the drug-carrier complex.
  • the second nucleotide strand hybridizes with at least one of the nucleotide strands of the drug-carrier complex.
  • the second nucleotide strand may be, as described above for the nucleotide strand of the drug-carrier complex, one or more nucleotides, single stranded, double stranded, DNA, RNA, naturally occurring or synthetic nucleotides.
  • the invention in another embodiment, relates to a method of forming a drug- carrier complex, comprising the steps of combining a drug with at least two nucleotide strands that hybridize with each other.
  • the drug and nucleotide may be added to a solution (e.g, water) either individually or together.
  • a drug and nucleotide are combined individually to form the drug-carrier complex, the drug may be added to the solution and then the nucleotide added to the solution or the nucleotide may be added to solution and then the drug added to the solution.
  • the drug and nucleotide may be added to the solution at the same time.
  • the drug and nucleotide may be added to the solution at the same time.
  • drug may be either insoluble or have relatively low solubility in water, such as a solubility of less than about one mg/liter at 25°C.
  • the dissolved drug-carrier complex is lyophilized. Lyophilization is also refened to as freeze-drying. Methods to lyophilize substances, which can be used to lyophilize the drug-carrier complexes of the invention, are well known in the art.
  • the solution of drug-carrier complex can be frozen (e.g., by placing in a liquid nitrogen or dry ice alcohol bath) and the frozen drug-carrier complex can be placed in a high vacuum.
  • the water in the form of ice
  • Another embodiment of the invention is a method of forming a drug-carrier composition, comprising the steps of combining a drug component and a nucleotide component.
  • the combined drug and nucleotide components are lyophilized to form the drug-carrier composition.
  • at least one of the drug components e.g., an antisense oligonucleotide, , anthracycline, distamycin
  • the nucleotide component e.g., single stranded DNA or RNA, double stranded DNA or RNA
  • the remaining components then may be added to the combined drug and nucleotide component.
  • a drug-carrier composition is formed by a method that includes lyophilizing a drug component, lyophilizing a nucleotide component and combining the lyophilized drug component and the lyophilized nucleotide component to form the drug-carrier composition.
  • the invention relates to a drug carrier that includes a double-stranded nucleotide (e.g., DNA or RNA) and a polymer component covalently bonded to at least one strand of the double stranded nucleotide.
  • the polymer component of the drug carrier has an aqueous solubility of at least one mg/liter at 25°C.
  • polymer generally refers to a molecule (e.g., protein, polyether, polyacetal, polysaccharide) formed by the union or bonding of chemically similar or chemically distinct units (e.g., monomers such as amino acids, glucose). Generally, “polymer” refers to a molecule comprising greater than about 30 units.
  • the polymers can be, for example, inorganic polymers such as siloxanes or polyphosphates and derivatives thereof. Alternatively, or additionally, the polymer can be organic.
  • Organic polymers can be natural organic polymers such as polysaccharides, starch, cellulose, pectin, inulin, agarose, chondroitinsulfate, heparin, dextrans, polypeptides (e.g., casein, albumin, globulin, keratin, insulin, polylysine) and derivatives thereof.
  • Organic polymers can be synthetic organic polymers such as polyacetals, polyacrylates, polyvinyl alcohol, polyvinylpynolidone, polyethylene glycol, polyesters, polyamides, polyamines and derivatives thereof.
  • Organic polymers may be semisynthetic organic polymers such as methycellulose, modified starches and derivatives thereof.
  • the polymer component of the drug carrier is a biocompatible polymer component.
  • biocompatible refers to a polymer that does not invoke an adverse reaction (e.g., immune response) from an organism (e.g., a mammal), a tissue culture or a collection of cells, or if the adverse reaction does not exceed an acceptable level.
  • the biocompatible polymer component is selected from the group consisting of a polysaccharide (e.g., poly -D-glucose, polysialic acid, dextran, chondroitinsulfate, starch), a polyether and a polyacetal (e.g., poly[hydroxymethylethylene hydroxymethylformal]).
  • the biocompatible polymer is cross-linked.
  • the polymer component of the drug carrier may be one or more chemically similar polymers (e.g., polysaccharide polymer:polysaccharide polymer, polypeptide polyme ⁇ polypeptide polymer) or chemically distinct polymers (e.g., a polysaccharide polymer and a polypeptide polymer; a polyether polymer and a polyacetal polymer; a polysaccharide polymer, a polypeptide polymer, and a polyether polymer).
  • the polymer component of the drug carrier includes at least one polymer covalently bonded to at least one strand of the nucleotide.
  • the polymer component includes at least two chemically similar or chemically distinct polymers.
  • An additional embodiment of the invention relates to a drug carrier, comprising a double-stranded nucleotide and an oligomer component covalently bonded to at least one strand of the double-stranded nucleotide.
  • oligomer refers to a molecule formed by the union or bonding of chemically similar or chemically distinct units (e.g., monomers such as amino acids, glucose, galactose), generally comprising less than about 30 units. Similar to polymers, oligomers can be, for example, inorganic oligomers or organic oligomers. Organic oligomers can be natural organic oligomers, synthetic oligomers or semisynthetic organic oligomers.
  • the oligomer component of the drug carrier may include at least one oligosaccharide, or at least one ohgopeptide, or a combination of both an oligosaccharide and an ohgopeptide.
  • the oligomer component includes at least one oligomer covalently bonded to at least one strand of the nucleotide.
  • the oligomer component includes at least two oligomers.
  • the oligomer component of the drug carrier includes at least two chemically distinct oligomers.
  • the oligomer component of the drug carrier includes at least two chemically similar oligomers.
  • the invention is a drug-carrier complex, that includes a single-stranded nucleotide, a polymer and a drug.
  • the drug is reversibly associated with the single-stranded nucleotide.
  • the polymer is associated with the single-stranded nucleotide. In another embodiment, the polymer is associated with the drug.
  • the invention is a drug-carrier complex, that includes a single-stranded nucleotide, an oligomer and a drug.
  • the oligomer is associated with the single-stranded nucleotide.
  • the oligomer is associated with the single-stranded nucleotide by a covalent association.
  • the oligomer can be associated with the drug.
  • the drug us reversibly associated with the single-stranded nucleotide.
  • the drug is reversibly associated with the oligomer.
  • the invention is a drug carrier that includes a single-stranded nucleotide and at least two polymers associated (e.g., reversibly, ineversibly) with the single-stranded nucleotide.
  • the chemical association e.g., reversible, ineversible
  • the chemical association e.g., reversible, ineversible between the polymer and the single- stranded nucleotide is a covalent bond.
  • the chemical association (e.g., reversible, ineversible) between the polymer and the single- stranded nucleotide is a noncovalent bond.
  • the invention is a drug carrier that includes a single- stranded nucleotide and at least two oligomers associated with the single-stranded nucleotide.
  • the chemical association between the oligomer and the single-stranded nucleotide is a covalent bond. In another embodiment, the chemical association between the oligomer and the single-stranded nucleotide is a noncovalent bond.
  • the invention relates to a drug-carrier composition
  • a drug-carrier composition comprising a nucleotide carrier component and a drug component.
  • the drug-carrier composition has a moisture content less than about 5% by weight.
  • An additional embodiment of the invention relates to a drug-carrier composition consisting essentially of a drug component and a nucleotide component, wherein other components, such as water or water vapor, essentially are absent.
  • the invention relates to a pharmaceutical formulation, that includes a nucleotide carrier component and a drug in reversible association with the nucleotide carrier component.
  • the reversible association of the drug and nucleotide carrier component is as described above for the drug and the nucleotide of a drug-carrier complex and may include, for example, van der Waals force, an electrostatic interaction, a hydrogen bond, an ionic bond, a hydrophobic interaction or a donor/acceptor bond.
  • the reversible association between the drug and the nucleotide carrier component of the pharmaceutical formulation may include at least one reversible association selected from the group consisting of a van der Waals force, an electrostatic interaction, a hydrogen bond, an ionic bond, a hydrophobic interaction and donor/acceptor bond.
  • the reversible association also may include intercalation between the drug and the nucleotide carrier component.
  • the reversible association between the drug and the nucleotide carrier component of the pharmaceutical formulation can further include a reversible covalent bond between the drug and the nucleotide carrier component.
  • the nucleotide carrier component of the pharmaceutical formulation is a polynucleotide carrier component. In another embodiment, the nucleotide carrier component of the pharmaceutical carrier component is an oligonucleotide carrier component.
  • the drugs that are used in the pharmaceutical formulation are as described above, for example, in drug-carrier complexes of the invention.
  • the drug in the pharmaceutical formulation may be an oligonucleotide drug (e.g., an oligonucleotide, an antisense oligonucleotide or a ribozyme).
  • the drug can include a component selected from the group consisting of an intercalator, a metal containing substance (e.g., platinum, cis-platinum, carboplatin, platinum, gold, silver), a minor groove binder or a major groove binder.
  • the drug includes at least one amino group.
  • the drug doxorubicin includes an amino group.
  • the drug of the pharmaceutical formulation is a protein or a peptide comprising two or more amino acids.
  • the amino acids of the protein or peptide drug can be naturally occurring L-amino acids, D-amino acids, nonnaturally occurring amino acids or synthetic amino acids such as gamma amino acids and cyclic amino acids.
  • the proteins can be post-translationally modified (e.g, glycosylated, myrisilated, acetylated).
  • the N-terminus amino group, the C-terminal carboxyl group, or one or more of the peptide bonds in the protein can be, for example, a non-ammo linkage.
  • the drug of the pharmaceutical formulation includes a diagnostic label.
  • diagnostic label when referring to a drug in the pharmaceutical formulation of the invention, refers to a detectable label incorporated into the drug. The label is used to determine the concentration of the drug, or a drug metabolite, in a certain sample, liquid, organ, tissue, combination of cells, single cell, cell organelle, or elsewhere.
  • Labels include, for example, a radionuclide, a fluorophore, a chromophore, a paramagnetic ion or moiety, a superparamagnetic nanoparticle, a barium or other heavy metal ion, a heavy metal (e.g., gold) particle, an iodine atom, an enzyme, or biotin.
  • the label can be detectable in vitro or in vivo, for example, by radioactivity measurement, gamma scintography, positron emission tomography, nuclear magnetic resonance spectroscopy, magnetic resonance imaging, fluorescence spectroscopy, photoimaging, X-ray (computed) tomography, electron microscopy, enzyme essay, or other respective methods.
  • the information on the label location or content can be used to determine the pathways of drug transfer and metabohzation.
  • the diagnostic label also can be used to confirm the presence of a disorder or a disease.
  • An additional embodiment of the invention relates to a method of delivering a drug to an organism, comprising administering a nucleotide carrier-drug complex to the organism.
  • the nucleotide carrier-drug complex includes a nucleotide carrier and a drug in reversible association with each other.
  • the reversible association of the drug and the nucleotide carrier component, used in the method of delivering a drug to an organism is selected from the group consisting of a van der Waals force, an electrostatic interaction, a hydrogen bond, an ionic bond, a hydrophobic interaction and donor/acceptor bond.
  • the reversible association of the drug and the nucleotide carrier component used in the method of delivering a drug to an organism is an intercalation.
  • the organism to which the drug is delivered using the method of the invention is selected from the group consisting of a mammal and a cell.
  • the mammal may be, for example, a primate (e.g., human, rhesus monkey), rodent (e.g., hamster, mouse, rat), ruminant (e.g., sheep, horse, cow) or domestic (e.g., cats, dogs) mammal.
  • the cell may be a procaryotic cell or eucaryotic cell.
  • the drug-carrier complex is administered into the organism by, for example, systemic or local administration. In another embodiment of the method, the drug-carrier complex is administered proximate to the organism. "Proximate to the organism,” as used herein, means near the organism. For example, if the organism is a cell, then the nucleotide carrier may be administered into the cell culture media.
  • the invention relates to a method of delivering a drug to a tissue culture or a combination of cells (e.g., tissue sample), comprising administering a nucleotide carrier-drug complex to a tissue culture.
  • the nucleotide carrier-drug complex includes a nucleotide carrier and a drug in reversible association with each other.
  • the reversible association of said drug and said nucleotide carrier is selected from the group consisting of a van der Waals force, an electrostatic interaction, a hydrogen bond, an ionic bond, a hydrophobic interaction and donor/acceptor bond.
  • the reversible association between the drug and the nucleotide carrier component in the method can be an intercalation.
  • the drug- carrier complex may be administered into the tissue culture or a combination of cells.
  • the nucleotide carrier can be administered proximate to the tissue culture or a combination of cells.
  • drugs of the nucleotide carrier-drug complex can be assessed to determine whether the drug inhibits cellular proliferation of tissue culture cells, or a pathogen such as: normal or cancer cells (e.g., melanoma, mammary adenocarcinoma cell line 293, tissue bioptates), viruses (e.g., HIV, Hepatitis C); bacteria, fungi and protozoa.
  • a pathogen such as: normal or cancer cells (e.g., melanoma, mammary adenocarcinoma cell line 293, tissue bioptates), viruses (e.g., HIV, Hepatitis C); bacteria, fungi and protozoa.
  • Another embodiment of the invention is a method of delivering a drug to an organism, comprising the step of administering a drug and a nucleotide carrier, which reversibly associates with the drug to form a nucleotide carrier-drug complex, to the organism.
  • the drug and the nucleotide carrier are administered simultaneously to the organism.
  • the drug and the nucleotide carrier are administered separately to the organism.
  • the drug may be administered first, followed by the nucleotide carrier, or the nucleotide carrier may be administered first, followed by the drug.
  • the invention relates to a method of delivering a drug to an organism, comprising forming a nucleotide carrier-drug complex that includes a drug and a nucleotide carrier in reversible association with the drug, and administering the nucleotide carrier-drug complex to the organism.
  • the invention in still another embodiment, relates to a method of delivering a drug to an organism, comprising administering to the organism a drug-carrier complex.
  • the drug-carrier complex includes a drug component and a carrier component in reversible association with each other.
  • the drug dissociates from the drug-carrier complex and reassociates with the carrier component.
  • the degree of association and dissociation can depend, for example, on the concentration of the drug and the carrier, and can be assessed using uv/vis spectroscopy, fluorescent spectroscopy, NMR or other suitable methods.
  • the carrier component is a nucleotide carrier component.
  • the nucleotide carrier component can be an oligonucleotide or a polynucleotide.
  • the nucleotide carrier component can be a single stranded nucleotide, a double stranded nucleotide, DNA, RNA, naturally occurring or synthetic nucleotides.
  • the drug-canier complex employed in the method of delivering a drug to an organism, is delivered to a combination of cells in said organism.
  • the combination of cells may be, for example, a cancer (e.g., breast cancer, brain cancer, prostate cancer, lung cancer), a pathogenic organism (e.g., bacteria, virus, fungal), or an organ (e.g., heart, kidney, lung, intestine, stomach).
  • the drug-carrier complex is administered to an organism and the drug-carrier complex dissociates near or within a combination of cells within the organism.
  • the drug-carrier complex is administered to a human organism and dissociates at or within cancer tissue within the human.
  • the drug-carrier complex may be administered to the organism at a point remote from the combination of cells of interest. Alternatively, or additionally, the drug-carrier complex may be administered to the organism at a point proximate to the combination of cells of interest. For example, if the combination of cells is lung cancer, administration of the drug-carrier complex via inhalation can be considered administering the drug at a point proximate to the combination of cells. Likewise, if the combination of cells is a small intestine cancer, administration of the drug-carrier complex into the peritoneum is considered to be an administration of the drug at a point proximate to the combination of cells.
  • the nucleotide carrier-drug complexes and pharmaceutical formulations of the invention may be administered systemically or locally, for example intravenously, intramuscularly, parenterally, orally, nasally, by inhalation, or by suppository.
  • the nucleotide carrier-drug complexes and pharmaceutical formulations of the invention may be administered in a single dose or in more than one dose over a period of time required to achieve a desired effect (e.g., delivery of a drug to a tumor to radiosensitize cancer cells or to decrease or halt cell proliferation).
  • the nucleotide carrier-drug complexes and pharmaceutical formulations of the invention can be admixed or combined with other pharmaceutical carriers or excipients such as sterile water, salt solutions (such as Ringer's solution), alcohols, or talc to facilitate administration to the organism, tissue culture or combination of cells.
  • the nucleotide carrier-drug complexes and pharmaceutical formulations of the invention can be sterilized and if desired, mixed with auxiliary substances, e.g., cryoprotectors, colorants or preservatives which do not deleteriously react with the nucleotide carrier-drug complexes and pharmaceutical formulations.
  • nucleotide carrier- drag complexes and pharmaceutical formulations of the invention in a specific case may vary according to the specific nucleotide carrier-drug complexes and pharmaceutical formulations being utilized, for example, the mode of administration and the age, weight and disease or disorder of the organism (e.g., a human).
  • the invention relates to a method for increasing aqueous solubility of a substance, comprising reversibly associating the substance with a nucleotide canier to form a water-soluble complex.
  • a nucleotide canier for example, several known bis-intercalators, such as doxorubicin, WP-631, and DMP 840 (Raghavan, K.S., et al, Pharm. Dev. Technol 37:3078-85 (1988), the teachings of which are hereby incorporated by reference in their entirety) have limited solubility at physiological pH.
  • aqueous solubility generally refers to the ability of a substance to blend (e.g., dissolve) with a water-based solution.
  • the water-based solution can be any solution that contains as one of its components water.
  • the water-based solution can be blood plasma or a physiologically buffered salt solution such as phosphate buffered saline or Ringer's solution.
  • the substance, without association with the nucleotide carrier is essentially insoluble in water.
  • the insolubility in water can render the substance inadequate or inefficient for administration to an organism.
  • increasing the aqueous solubility employing the methods of the invention provides an improved method of delivering drugs to cells, organisms (e.g., mammals) and tissue cultures to treat and study the mechanism of disease and increases the number of compounds that can be used as drugs.
  • the invention relates to a targeted carrier, comprising a nucleotide, a polymer component associated with the nucleotide and a ligand associated with the nucleotide or the polymer component, and associable with a cell or tissue marker.
  • the association between the ligand and the nucleotide or polymer component of the targeted carrier can be a reversible (e.g., a van der Waals force, an electrostatic interaction, a hydrogen bond, an ionic bond, a hydrophobic interaction and donor/acceptor bond), a nonreversible association or a covalent bond.
  • the cell or tissue marker with which the ligand of the targeted carrier associates can be, selected, for example, from the group consisting of proteins, polysaccharides, polypeptides, carbohydrates and lipids.
  • the terms “proteins,” “polysaccharides,” “polypeptides,” “carbohydrates,” and “lipids” are intended to also refer to related compounds, or derivatives, such as glycoproteins, glycolipids, lipopolysaccharides, proteoglycans, lipoproteins, lipid- protein complexes, nucleosomes, and lipoteichoic acids.
  • the cell or tissue marker can be a cell surface receptor such as transmembrane receptor (e.g., G- protein coupled receptors, tyrosine kinase receptors, growth factor receptors).
  • the ligand of the targeted carrier may be selected to specifically target the targeted carrier to a particular cell or tissue to, for example, deliver a drug to treat a disease condition or to compensate for a deficiency.
  • Tissue markers can be divided, for example, in two major groups.
  • One group can comprise of molecules expressed exclusively or almost exclusively on the surfaces of pathological cells or in the pathological extracellular matrix, or in certain combinations of cells (cell types, cell classes, organs, or tissues).
  • Another group can comprise molecules that are not unique to the pathological sites, but are overexpressed in the pathological sites.
  • suitable ligands that associate with the cell or tissue marker are platelet-derived growth factor (PDGF), macrophage colony-stimulating factor and epidermal growth factor.
  • the typical representatives of the first group are asialofetuin receptors (hepatocytes), viral antigens (cells infected by herpes or other viruses), scavenger receptors (macrophages), HER-2/neu (some breast cancer types).
  • the second group includes markers typical for several types of inflammation and cancer, for example, cytokine receptors, receptors of growth factors, surface glycolipids and glycoproteins, integrins, selectins, etc.
  • malignant epithelial cells in primary human lung carcinomas coexpress in vivo PDGF (ligand) and PDGF receptor (cell or tissue marker)
  • PDGF ligand
  • PDGF receptor cell or tissue marker
  • macrophage colony-stimulating factor (ligand) and its receptor are expressed in ovarian and endometrial carcinomas
  • coexpression of HER-2/neu and the epidermal growth factor receptor (cell or tissue marker) has been observed in 65% of epithelial ovarian cancers and in a limited number of normal tissue from a fraction of donors (Bast, R.C., Jr., et al, Cancer, 77:1597-601 (1993), the teachings
  • a targeted carrier that includes a nucleotide and a polymer.
  • the polymer component of the targeted carrier is a ligand associable with a cell or tissue marker.
  • the cell or tissue marker is selected from the group consisting of proteins, polypeptides, carbohydrates, lipids and nucleotides and derivatives thereof.
  • a cell or tissue marker can also be a glycolipids, glycoproteins, glycopeptides, transmembrane proteins, glycoproteins and proteoglycans of the extracellular matrix and other molecules present in tissues and exposed to the extracellular environment. This also includes intracellular components exposed to the extracellular environment in disease (e.g., nucleosomes).
  • the invention is a targeted drug-carrier complex, comprising a nucleotide, a drug reversibly associated with the nucleotide and a targeting component.
  • the targeting component is associated with the nucleotide or the drug.
  • the targeting component includes a ligand associable with a cell or tissue marker.
  • the ligand can be, for example, associable with the cell or tissue marker by covalent, noncovalent or reversible associations
  • the cell or tissue marker is selected from the group consisting of proteins, polypeptides, carbohydrates, lipids and nucleotides.
  • the drug is reversibly associated with the nucleotide.
  • the targeting component is associated either with the drug or with the nucleotide.
  • the invention is a targeted drug-carrier complex that includes a nucleotide, a drug reversibly associated with the nucleotide, a polymer component and a targeting component.
  • the polymer component is associated with the nucleotide or the drug.
  • the targeting component is associated with the nucleotide, the drug or the polymer.
  • the association between the targeting component and the drug or polymer can be, for example, a covalent bond, noncovalent bond or a reversible association.
  • the targeting component includes a ligand associable with a cell or tissue marker and a drug.
  • the association between the targeting component and the ligand can be, for example, a covalent bond, noncovalent bond or a reversible association.
  • the cell or tissue marker is selected from the group consisting of proteins, polypeptides, carbohydrates, lipids and nucleotides.
  • the invention relates to a drug delivery system, comprising a matrix, a nucleotide associated with or entrapped within the matrix, and a drug in reversible association (e.g., a van der Waals force, an electrostatic interaction, a hydrogen bond, an ionic bond, a hydrophobic interaction and donor/acceptor bond) with the nucleotide ( Figure 4).
  • the matrix of the drug delivery system is a gel, a film or a particle. The matrix provides a structural foundation of the drug delivery system.
  • the matrix can minimize any adverse reaction an organism, cell or tissue culture may have to the drug delivery system.
  • the matrix is biocompatible.
  • Matrix materials generally are selected in accordance with the method of administration of the drug release system. In topical systems, such as gels, films, patches and other systems for external application, hydrophilic gels frequently are used as matrix materials. Gels are made, for example, of biocompatible polymers such as collagen, fibrin, polyvinylpynolidone, polyvinyl alcohol, polyethyleneglycol, polypropyleneglycol, polyacrylates, or combinations thereof. Other systems, such as liniments, include emulsions, suspensions, and liposomal preparations, sometimes in mixtures with each other or entrapped within a gel.
  • Implantable drug delivery systems are described below as implants.
  • the matrix of the drug release system remains stable for as long as the drug release system remains functional, e.g., for as long as the drug is being released at a desirable rate.
  • the structural matrix of the drug release system would disintegrate rapidly. In an embodiment of the invention, this effect is achieved by matrix stabilization by the drug.
  • the matrix of the drug release system 66 (e.g., a hydrophilic polymer gel in Figure 4) is reversibly crosslinked via cross-hybridization of short ohgonucleotides 70 chemically associated (e.g., covalently bound) with the matrix material 68 ( Figure 4).
  • melting temperature of the double-stranded links formed as a result of hybridization is near or below normal body temperature. Association of a drug 72 with the double-stranded oligonucleotide stabilizes the latter and increases the melting temperature, making the gel stable at body temperature ( Figure 4).
  • the invention relates to an implant, comprising an implant matrix, a nucleotide associated with or entrapped within the matrix, and a drug in reversible association (e.g., a van der Waals force, an electrostatic interaction, a hydrogen bond, an ionic bond, a hydrophobic interaction and donor/acceptor bond) with the nucleotide.
  • a drug in reversible association e.g., a van der Waals force, an electrostatic interaction, a hydrogen bond, an ionic bond, a hydrophobic interaction and donor/acceptor bond
  • Implants capable of sustained drug release can be useful, for example, for prolonged systemic delivery of a drug after a single administration, for drug delivery to local lymph nodes draining the implantation site, or for postoperative local wound treatment.
  • the implant can be made, for example, of a solid material or a gel, and can be made as a single block (tablet, film) or consist of multiple particles.
  • the structural foundation of the implant can be engineered as a sponge, foam, fabric, thread, or otherwise.
  • the implant can be of any size and shape suitable for implantation. Particulate, thread or film implants can be more suitable for minimally invasive methods (such as implantation through a needle or other surgical tool), whereas other types of implants may be more suitable for conventional surgery.
  • the implants should perform precise biological functions, e.g., mediated by controlled release of biologically active compounds and/or direct structural and functional support of tissues and/or cell cultures, which can be performed best by macromolecular or supramolecular matrices comprising specialized functional domains. Matrices should be stable and biologically inert in vivo for a predetermined period of time, and completely degradable after their function has been completed. The biodegradation should not result in producing any toxic products nor polymer deposition in draining lymph nodes.
  • implant matrix materials are: silicone, copolymers of lactic and glycolic acids, agarose, porous metals (e.g., titanium), coral matrix, acrylates.
  • Other possible materials include polyethyleneglycol, polyacetals, polysaccharides, denatured or crosslinked proteins (e.g., albumin, gelatin) or natural proteins (e.g., collagen, fibrin).
  • the implant can be designed to release a certain dose of the drug over a particular period of time.
  • the matrix is a material reversibly cross-linked with a nucleotide/nucleotide association.
  • the release of the drug from the implant destabilizes the implant matrix.
  • EXAMPLE 1 HYBRIDIZATION OF SINGLE-STRANDED OLIGONUCLEOTIDES.
  • Two custom-synthesized single stranded ohgonucleotides 5' AAA TCT CCC AGC GTG CGC CAT AA 3' (SEQ ID NO: 1) and 5' tt AtG GCG CAC GCt GGG AGA ttt 3' (SEQ ID NO: 2), where t is an amino modified T, were purchased from a commercial source.
  • the resultant solution (total volume 22 ⁇ l) was transfened to a capped 1 ml vial, and the vial was heated to a 95°C in a 100 ml water bath for 10 minutes. Then the bath was allowed to cool down to 25°C.
  • the resultant product a double-stranded oligonucleotide, was purified by size exclusion HPLC in water and lyophilized. Yield: 91%.
  • EXAMPLE 2 FORMATION AND ISOLATION OF CARRIER-DRUG COMPLEX.
  • oligonucleotide with arbitrarily chosen 18-base sequence 5'CGT CGA CGT CGA ATA TAC GC (SEQ ID NO: 3), and a complementary 5'-amino modified oligonucleotide 5'GC GTA TAT TCG ACG TCG ACG (SEQ ID NO: 4) were purchased from a commercial vendor.
  • SEQ ID NO: 3 5'CGT CGA CGT CGA ATA TAC GC
  • SEQ ID NO: 4 a complementary 5'-amino modified oligonucleotide 5'GC GTA TAT TCG ACG TCG ACG (SEQ ID NO: 4) were purchased from a commercial vendor.
  • the single-stranded ohgonucleotides were hybridized and lyophilized as described in Example 1.
  • the resultant double stranded oligonucleotide was stable at ambient (25°C) and body temperature (37° C).
  • the double-stranded oligonucleotide was dissolved in water at 2 mg/ml without pH adjustment.
  • the solutions 0.1 ml each, were mixed.
  • 0.3 ml of PBS, pH : 7, were added, and the resultant solution was incubated at ambient temperature for 10 minutes.
  • the reaction mixture was purified by gel chromatography on Sephadex G-25 in water. Doxorubicin elution was monitored photometrically at 470 nm. Essentially all doxorubicin was found in the oligonucleotide fraction.
  • EXAMPLE 3 DRUG-CARRIER ADDUCT WITH HIGH DRUG CONTENT Drug-carrier adduct with a high drug content was prepared essentially as described in Example 2, using the same double stranded oligonucleotide and doxorubicimoligonucleotide ratio 1 :5 (w/w), which conesponds to approximately one doxorubicin molecule per four base pairs.
  • the resultant adduct was purified by gel chromatography (PD-10 column, water) and lyophilized. Yield: 98 ⁇ 2%.
  • adducts obtained as described in Examples 2 and 3 were used in a lyophilized form.
  • Doxorubicin powder (Sigma Chemical Co., St. Louis, MO) and lyophilized "Doxorubicin for injection" were used as control preparations.
  • oligonucleotide-doxorubicin adduct solutions were filtered through 0.22 mm PTFE membrane filters; doxorubicin was recovered in the filtrates with at least 98% yield (by adsorption at 470 nm). Filtration of the suspensions of the control preparations through 0.22 mm PTFE membrane resulted in the recovery of less than 5% of doxorubicin in the filtrate; the rest was retained by the filter.
  • Doxorubicin solution 0.1 ml of a 0.1 mg/ml solution, was applied to a short column packed with Sephadex G-25 (PD-10, Pharmacia). Formation of doxorubicin-Sephadex adduct was detected by formation of a characteristically colored red layer. Elution with water resulted in 0% doxorubicin recovery in the first 10 ml, with subsequent slow elution. Analogous experiment with doxorubicin adducts with model carrier (Examples 2, 3) resulted in complete doxorubicin elution in a 2.5 ml fraction (2.5 to 5.0 ml).
  • doxorubicin was adsorbed on the PD-10 column as described above, and the column was washed with 10 ml H 2 O. Then the double-stranded oligonucleotide of Example 2, 0.1 ml of a 1 mg/ml solution, was passed through the same column. Doxorubicin adsorbed on Sephadex G-25 was completely desorbed from the column, and eluted within the oligonucleotide fraction (2.5 to 5.0 ml).
  • EXAMPLE 6 ELECTROPHORESIS OF DOXORUBICIN ADDUCTS WITH MODEL DRUG-CARRIER COMPLEXES
  • Anti-fluorescein IgG was added at 10-fold concentration to ensure pseudo first order conditions. Fluorescein - antibody interaction was registered by quenching of the fluorescein fluorescence by the antibody. Fluorescence was registered at 515 nm (excitation at 490 nm). The kinetics of fluorescence quenching depended on the position of the fluorescein moiety relative to the polymer chain. Polymer chains positioned at 16-20 bases did not attenuate fluorescein-antibody interaction, whereas polymer chains positioned within 2 and 9 bases did decrease the kinetic constant by 20% (2 kDa polymer) to 80% (20 kDa polymer).
  • the degree of steric protection of the nucleotide can be optimized via optimization of the distance between polymer chains (number of chains per base pair) and of the molecular weight of the polymer. Based on the data described in the following examples, a 50% to 80% hindrance of the carrier core (as measured by protein access kinetics) can be sufficient to prolong canier circulation by several hours.
  • EXAMPLE 8 MODEL STERICALLY PROTECTED (POLYMER-MODIFIED) CARRIERS
  • the latter polymer was prepared via oxidation of Dextran B512 with 1.8 periodate molecules per carbohydrate ring, with subsequent borohydride reduction and second periodate oxidation of the resultant glycol groups.
  • Aldehydo-PHF 50-fold excess was reacted with the amino modified carrier core in the presence of cyanoborohydride (1 mole per mole aldehyde) overnight at ambient temperature.
  • cyanoborohydride (1 mole per mole aldehyde
  • EXAMPLE 9 DRUG-CARRIER COMPLEX CONJUGATION WITH MODEL ANTIBODY
  • a PHF-modified carrier analogous in structure to the carrier of Example 8 but comprising glycol groups in the polymer chains, was prepared using a modified technique.
  • the amino modified carrier was modified with 10 kDa poly(hydroxymethylethylene hydroxymethylformal) (PHF) containing 10% aldehyde groups and 10% glycol groups.
  • PHF poly(hydroxymethylethylene hydroxymethylformal)
  • the conjugate was separated from the unreacted IgG by HPLC (yield by IgG absorption at 280 nm: 22 ⁇ 9%).
  • the calculated amount of active IgG in the carrier was 1 ⁇ 0.3% w/w.
  • the product was isolated by HPLC.
  • EXAMPLE 11 DRUG-CARRIER ADDUCT MODIFICATION WITH PHF VIA (AMINOOXY)DOXORUBICIN
  • AHD N-(3-aminooxy-2-hydroxypropyl)-doxorubicin
  • Adduct modification with polymer chains was detected by decrease in the elution time of the adduct (6.2 min vs. 8.5 min for unmodified adduct).
  • polymer incubation with analogous unloaded carrier the elution time of the latter remained unchanged.
  • EXAMPLE 12 CARRIER LOADING WITH RADIOLABELED DOXORUBICIN
  • a sterically protected drug-carrier complex 0 of Example 8 was loaded with a bis-intercalator WP-631 synthesized via crosslinking of two daunorubicin molecules with ⁇ -Dibromo-p-xylene as described in the literature (Chaires, J.B., et al, J. Med. Chem. 40:261-6 (1997), the teachings of which are hereby incorporated by reference in their entirety).
  • Bis-intercalator content was one molecule per 6 base pairs of the nucleotide core.
  • unloaded carrier was injected at 100 mg/kg within 15 minutes after the administration of 30 mg/kg of bis-doxorubicin.
  • Unloaded carriers (Example 8) were labeled with tritium ( 3 H). Carriers were oxidized with periodate to produce aldehyde groups on the 3'-ribonucleotide base present in the structure, and reacted with [ 3 H] borohydride to introduce tritium into carrier core structure. Carriers were purified on Sephadex G-25 and injected at 1 mg/kg into normal outbred mice (males, 30 g) via tail vein. Blood samples were collected at different time points and counted. The blood half-lives were ca. 30 min and 10 hours for unprotected and PHF-protected carriers, respectively. This example shows that circulation time of the nucleotide-based carriers can be optimized by steric protection, and long-circulating carriers can be prepared via modification with hydrophilic polymers.
  • doxorubicin adduct with PHF-protected carrier resulted in twice higher label content in skeletal muscle, 6 ⁇ 2 vs. 2 ⁇ 0.4 % dose/g (mean ⁇ standard deviation).
  • Doxorubicin adduct with DNA showed increased label distribution to lung (7 ⁇ 3.1% vs. A.6 ⁇ 1.6% dose/g) and liver (12 ⁇ 6 vs. 3 ⁇ 0.6 % dose/g).
  • EXAMPLE 16 MODEL pCMV-BASED CARRIER
  • a plasmid solution in PBS, pH 7.5, 0.1 ml, containing 10 mg/ml pCMN (Promega), was mixed with 10 ml of 1 mg/ml solution of bis-doxorubicin (described in Example 13). After a 1 hr incubation, the adduct was isolated by SEC HPLC (BioRad BioSil 125 column). Bis-doxorubicin association with the plasmid was detected by appearance of absorption at 470 nm in the excluded volume.
  • EXAMPLE 18 OLIGONUCLEOTIDE CONJUGATION WITH POLYLYSINE 5'-amino modified oligonucleotide with sequence
  • the product was purified by SEC on Sephadex G-25 (PD-10 column).
  • Polylysine hydrobromide, 20 kDa, 0.1 ml of 2 mg/ml solution, was mixed with 0.1 ml of a 1 mg/ml of the oligonucleotide solution in PBS, pH 6.
  • EDC ethyl-(N-dimethylaminopropyl)carbodiimide
  • N-hydroxysuccinimide ester of terminal-carboxypropylthio-PHF was prepared via PHF reaction with mercaptopropionic acid with subsequent modification with N-hydroxysuccinimide in the presence of dicyclohexylcarbodiimide.
  • the conjugate synthesized in Example 18 (ca. 0.1 mg in
  • N-hydroxysuccinimide ester of terminal-carboxypropylthio-PHF 10 mg was dissolved in 0.1 ml DMSO. The solutions were mixed and incubated at room temperature overnight. The product was isolated by size exclusion HPLC (BioSil

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Abstract

La présente invention concerne des complexes vecteurs de médicament, des vecteurs de médicament, des préparations pharmaceutiques, et des techniques d'apport de ces médicaments dans un organisme ou dans une culture de tissus, des techniques permettant d'augmenter la solubilité d'une substance, des vecteurs ciblés, des systèmes d'apport de médicament et des implants. Les compositions et les techniques de cette invention comprennent la préparation de complexes dont les associations entre des nucléotides et des médicaments sont réversibles. Ces compositions et ces techniques peuvent être utilisées pour cibler des médicaments sur des cellules, des organismes ou des combinaisons de cellules à traiter, pour étudier les mécanismes sous-jacents de maladies, et pour tester des médicaments candidats.
EP00955415A 1999-08-09 2000-08-09 Complexes vecteurs de medicament et technique d'utilisation de ceux-ci Withdrawn EP1206285A2 (fr)

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Families Citing this family (24)

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US8106098B2 (en) 1999-08-09 2012-01-31 The General Hospital Corporation Protein conjugates with a water-soluble biocompatible, biodegradable polymer
ATE410459T1 (de) 2002-01-14 2008-10-15 Gen Hospital Corp Bioabbaubare polyketale, verfahren zu ihrer herstellung sowie ihre verwendung
US8030459B2 (en) 2002-07-19 2011-10-04 The General Hospital Corporation Oxime conjugates and methods for their formation and use
EP1667726B1 (fr) 2003-09-05 2011-05-04 The General Hospital Corporation Conjugués comprenant un residu polyacétal et un medicament comme système de libération
AU2008268432B2 (en) 2007-06-25 2015-01-15 Endocyte, Inc. Conjugates containing hydrophilic spacer linkers
AR074584A1 (es) 2008-12-10 2011-01-26 Mersana Therapeutics Inc Formulaciones farmaceuticas de conjugados camtotecina-polimero biocompatibles biodegradables
CN105214143A (zh) 2009-04-28 2016-01-06 苏尔莫迪克斯公司 用于递送生物活性剂的装置和方法
US9861727B2 (en) 2011-05-20 2018-01-09 Surmodics, Inc. Delivery of hydrophobic active agent particles
US9757497B2 (en) * 2011-05-20 2017-09-12 Surmodics, Inc. Delivery of coated hydrophobic active agent particles
US10213529B2 (en) 2011-05-20 2019-02-26 Surmodics, Inc. Delivery of coated hydrophobic active agent particles
BR112013031819B1 (pt) 2011-06-10 2022-05-03 Mersana Therapeutics, Inc Suporte polimérico, composição farmacêutica, composto, e, uso do suporte
US8815226B2 (en) 2011-06-10 2014-08-26 Mersana Therapeutics, Inc. Protein-polymer-drug conjugates
JP5378469B2 (ja) * 2011-08-11 2013-12-25 学校法人 日本歯科大学 医療用薬剤
US8785569B2 (en) 2011-11-22 2014-07-22 Original Biomedicals Co., Ltd. Drug carrier with chelating complex micelles and the application thereof
US11246963B2 (en) 2012-11-05 2022-02-15 Surmodics, Inc. Compositions and methods for delivery of hydrophobic active agents
JP6438406B2 (ja) 2012-11-05 2018-12-12 サーモディクス，インコーポレイテッド 疎水性生理活性物質を送達するための組成物および方法
JP6420331B2 (ja) 2013-10-11 2018-11-07 メルサナ セラピューティクス，インコーポレイティド タンパク質−高分子−薬剤コンジュゲート
MX2016004659A (es) 2013-10-11 2017-08-02 Asana Biosciences Llc Conjugados proteina-polimero-farmaco.
EP3268046A4 (fr) * 2015-03-13 2018-11-21 Endocyte, Inc. Conjugués pour le traitement de maladies
US11474106B2 (en) 2015-07-08 2022-10-18 Lawrence Livermore National Security, Llc Methods for cytotoxic chemotherapy-based predictive assays
CA3039040A1 (fr) * 2016-10-03 2018-04-12 Eos Biosciences, Inc. Complexes therapeutiques comprenant un medicament a petite molecule complexe et un arn fonctionnel et vehicule d'administration nanoparticulaire
US10898446B2 (en) 2016-12-20 2021-01-26 Surmodics, Inc. Delivery of hydrophobic active agents from hydrophilic polyether block amide copolymer surfaces
WO2018132766A1 (fr) * 2017-01-12 2018-07-19 The Regents Of The University Of California Dosages prédictifs basés sur une chimiothérapie cytotoxique pour la leucémie myéloïde aiguë
WO2021118927A1 (fr) * 2019-12-13 2021-06-17 Insideoutbio, Inc. Méthodes et compositions pour l'administration ciblée d'agents thérapeutiques par l'acide nucléique

Citations (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
GB1440626A (fr) * 1973-05-02 1976-06-23 Farmaceutici Italia

Family Cites Families (5)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
DE3568583D1 (en) * 1985-08-29 1989-04-13 Berol Kemi Ab A carrier with an immobilised, biologically active substance, a method for its preparation, and the use thereof
US6610841B1 (en) * 1997-12-18 2003-08-26 Gilead Sciences, Inc. Nucleotide-based prodrugs
US5811510A (en) * 1995-04-14 1998-09-22 General Hospital Corporation Biodegradable polyacetal polymers and methods for their formation and use
AU3243300A (en) * 1999-02-23 2000-09-14 Isis Pharmaceuticals, Inc. Multiparticulate formulation
WO2000078285A1 (fr) * 1999-06-18 2000-12-28 University Of Medicine And Dentistry Of New Jersey Liberation regulee d'agents therapeutiques au moyen du piegeage in situ par reticulation de matrice

Patent Citations (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
GB1440626A (fr) * 1973-05-02 1976-06-23 Farmaceutici Italia

Non-Patent Citations (7)

* Cited by examiner, † Cited by third party
Title
HERSCHLAG ET AL: "An RNA chaperone activity of non-specific RNA binding proteins in hammerhead ribozyme catalysis", THE EMBO JOURNAL, vol. 13, 1994, pages 2913 - 2924, XP000567895 *
HUFF; KREUZER: "Evidence for a common mechanism of action for antitumor and antibacterial agents that inhibit type II DNA topoisomerases", THE JOURNAL OF BIOLOGICAL CHEMISTRY, vol. 256, 1990, pages 20496 - 20505 *
LIPSCOMB ET AL: "Water ring structure at DNA interfaces: hydration and dynamics of DNA-anthracycline conplexes", BIOCHEMISTRY, vol. 33, 1994, pages 3649 - 3659 *
PULLMAN: "Sequence specificty in the binding of anti-tumour anthracyclines to DNA: a success of theory", ANTI-CANCER DRGU DESIGN, vol. 7, 1991, pages 95 - 105 *
See also references of WO0110468A3 *
YANG; WANG: "Structural studies of atom-specific anticancer drugs acting on DNA", PHARMACOLOGY AND THERAPEUTICS, vol. 83, 1999, pages 181 - 215 *
ZIEGLER ET AL: "induction of apoptosis in small-cell lung cancer cells by an antisense oligodeoxynucleotide targeting the Bcl-2 coding sequence", JOURNAL OF THE NATIONAL CERCER INSTITUTE, vol. 89, 1997, pages 1027 - 1036, XP002080869, DOI: doi:10.1093/jnci/89.14.1027 *

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