[go: up one dir, main page]

WO1996004001A1 - Complexes biomoleculaires diriges - Google Patents

Complexes biomoleculaires diriges Download PDF

Info

Publication number
WO1996004001A1
WO1996004001A1 PCT/US1995/009870 US9509870W WO9604001A1 WO 1996004001 A1 WO1996004001 A1 WO 1996004001A1 US 9509870 W US9509870 W US 9509870W WO 9604001 A1 WO9604001 A1 WO 9604001A1
Authority
WO
WIPO (PCT)
Prior art keywords
complex
poly
group
alpha
derivatives
Prior art date
Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
Ceased
Application number
PCT/US1995/009870
Other languages
English (en)
Inventor
Robert Katz
Maria Tomoaia-Cotisel
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.)
Molecular/Structural BioTechnologies Inc
Original Assignee
Molecular/Structural BioTechnologies Inc
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
Priority claimed from US08/286,327 external-priority patent/US5716614A/en
Application filed by Molecular/Structural BioTechnologies Inc filed Critical Molecular/Structural BioTechnologies Inc
Priority to EP95929378A priority Critical patent/EP0952841A4/fr
Priority to AU32755/95A priority patent/AU3275595A/en
Publication of WO1996004001A1 publication Critical patent/WO1996004001A1/fr
Anticipated expiration legal-status Critical
Ceased legal-status Critical Current

Links

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
    • A61K47/62—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 a protein, peptide or polyamino acid
    • A61K47/64—Drug-peptide, drug-protein or drug-polyamino acid conjugates, i.e. the modifying agent being a peptide, protein or polyamino acid which is covalently bonded or complexed to a therapeutically active agent
    • A61K47/645—Polycationic or polyanionic oligopeptides, polypeptides or polyamino acids, e.g. polylysine, polyarginine, polyglutamic acid or peptide TAT
    • 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
    • A61K47/542—Carboxylic acids, e.g. a fatty acid or an amino acid
    • 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
    • A61K47/62—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 a protein, peptide or polyamino acid
    • A61K47/64—Drug-peptide, drug-protein or drug-polyamino acid conjugates, i.e. the modifying agent being a peptide, protein or polyamino acid which is covalently bonded or complexed to a therapeutically active agent
    • A61K47/6415—Toxins or lectins, e.g. clostridial toxins or Pseudomonas exotoxins

Definitions

  • the present invention relates to site-specific biomolecular lipophilic complexes, also called conjugates or transport vectors.
  • These complexes comprise an omega-3 fatty acid and derivatives thereof, and a therapeutic, prophylactic, diagnostic or a research agent.
  • said complex is conjugated with cationic macromolecular carriers to enhance its passage from the blood circulation to the brain.
  • the resultant conjugates are specific for sustained release delivery of said agents to the central nervous system (CNS), particularly to the neurons of the cerebral cortex, hippocampus and corpus striatum (e.g., cholinergic and adrenergic neurons) and to the glial tissue (neuroglial cells, including, astrocytes and macroglial cells) in animals and humans.
  • CNS central nervous system
  • targeting or internalizing moieties such as neurotoxins and fragments thereof, are attached to said complex to facilitate its uptake by the target brain cells.
  • compositions containing poly-unsaturated site-specific complexes/conjugates for the treatment of a wide variety of diseases causing to severe deterioration of the central nervous system, e.g., dementias, neurodegenerative disorders, neurological diseases, malignant brain tumors, inborn errors of metabolism (i.e., lysosomal storage disorders), and the like.
  • the purpose of this invention is to present a method that improves the transport and delivery characteristics of an agent molecule to a desired location, in the central nervous system,thus increasing its bioavailability.
  • agent as used herein relates to therapeutic, prophylactic and diagnostic compounds. These compounds are biologically active with beneficial effects in both animals and humans. Agents include lysosomal enzymes such as ceramidase, glucocerebrosidase, beta-galactosidase, beta-hexosaminidase A, beta- hexosaminidase A & B, galactosylceramidase, arylsulfatase A, sphingomyelinase, alpha-galactosidase B, aspartylglycosaminidase, alpha- L-fucosidase, iduronate sulfatase, alpha-L-iduronidase, glcNAc-6-sulfatase, beta-glucuronidase, their recombinant analogs and their derivatives.
  • serum proteins namely immunoglobulins, interleukins, interferons, hormones, such as insulin, parathyroid hormone, pigmentary hormone, thyroid-stimulating hormones, tissue plasminogen activator, nerve growth factors, peptidases or proteases, nucleic acids and derivatives thereof, nucleotides, oligonucleotides, antisense oligonucleotide analogs, genes, transfected cells, biological vectors, cloning vectors and expression vectors.
  • hormones such as insulin, parathyroid hormone, pigmentary hormone, thyroid-stimulating hormones, tissue plasminogen activator, nerve growth factors, peptidases or proteases, nucleic acids and derivatives thereof, nucleotides, oligonucleotides, antisense oligonucleotide analogs, genes, transfected cells, biological vectors, cloning vectors and expression vectors.
  • Neurotoxins or their non-toxic peptide fragments, diagnostic and research reagents are also included.
  • BBB blood-brain barrier
  • This layer separates two fluid-containing compartments: the blood plasma (BP) and extracellular fluid (ECF) of the brain parenchyma, and is surrounded by astroglial cells of the brain.
  • BP blood plasma
  • ECF extracellular fluid
  • One of the main functions of the BBB is to regulate the transfer of components between the BP and the ECF.
  • the BBB limits free passage of most agent molecules from the blood to the brain cells.
  • large molecules of high polarity such as peptides, proteins, (e.g., enzymes, growth factors and their conjugates, oligonucleotides, genetic vectors and others) do not cross the BBB. Therefore poor agent delivery to the CNS limits the applicability of such macromolecules for the treatment of neurodegenerative disorders and neurological diseases.
  • Drug delivery to the central nervous system through the cerebrospinal fluid is achieved by means of a subdurally implantable device named after its inventor the "Ommaya reservoir ".
  • the reservoir is used mostly for localized post-operative delivery of chemotherapeutic agents in cancers.
  • the drug is injected into the device and subsequently released into the cerebrospinal fluid surrounding the brain. It can be directed toward specific areas of exposed brain tissue which then adsorb the drug. This adsorption is limited since the drug does not travel freely.
  • a modified device developed by Ayub Ommaya whereby the reservoir is implanted in the abdominal cavity and the injected drug is transported by cerebrospinal fluid (taken from and returned to the spine) all the way to the ventricular space of the brain, is used for agent administration.
  • omega-3 derivatization site-specific biomolecular complexes of this invention are overcoming the limited adsorption and movement of the agent through brain tissue.
  • Prior art is silent on agents capable of site-directed penetration of brain tissue from the cere
  • Diffusion of macromolecules to various areas of the brain by convection- enhanced delivery is another method of administration circumventing the BBB.
  • This method consists of: a) Creating a pressure gradient during interstitial infusion into white matter to generate increased flow through the brain interstitium (convection supplementing simple diffusion); b) Maintaining the pressure gradient over a lengthy period of time (24 hours to 48 hours) to allow radial penetration of the migrating compounds (such as: neurotrophic factors, antibodies, growth factors, genetic vectors, enzymes, etc.) into the gray matter; and c) Increasing drug concentrations by orders of magnitude over systemic levels.
  • the migrating compounds such as: neurotrophic factors, antibodies, growth factors, genetic vectors, enzymes, etc.
  • the site-specific biomolecular complexes of this invention are instrumental in delivering the agent to neuronal or glial cells, as needed, and be retained by these cells. Moreover, the site-specific complexes containing neuronal targeting or internalization moieties are capable of penetrating the neuronal membrane and internalizing the agent. Prior art is silent on such site-specific delivery of macromolecular agents to specific sites in the central nervous system such as cortical and hippocampal neurons.
  • Another strategy to improve agent delivery to the CNS is by increasing the agent absorption (adsorption and transport) through the BBB and their uptake by the cells [Broadwell, Acta Neuropathol., 79: 117-128 (1989); Pardridge et al., J. Pharmacol. Experim. Therapeutics, 255(2): 893-899 (1990); Banks et al., Progress in Brain Research, 91: 139-148 (1992); Pardridge, Fuel Homeostasis and the Nervous System, Edited by Vranic et al., Plenum Press, New York, 43-53 (1991)].
  • the passage of agents through the BBB to the brain can be enhanced by improving either the permeability of the agent itself or by altering the characteristics of the BBB.
  • the passage of the agent can be facilitated by increasing its lipid solubility through chemical modification, and/or by its coupling to a cationic carrier, or still by its covalent coupling to a peptide vector capable of transporting the agent through the BBB.
  • Peptide transport vectors are also known as BBB permeabilizer compounds [U.S. Pat. No. 5,268,164].
  • Another major objective of this invention is to synthesize a site- specific macromolecule with lipophilic properties.
  • the resulting complex or conjugate which comprises an agent, one or more lipophilic moieties and a directing moiety, is targeted to specific cells in the brain.
  • the lipophilic moiety is either alpha- linolenic acid (ALA), eicosapentaenoic acid (EPA) or docosahexaenoic acid (DHA) or derivatives thereof (e.g.
  • alpha-linolenoyl-aminoethanol N-eicosapentaenoyl- aminoethanol, N-docosahexaenoyl-amino ethanol,or alpha linolenoyl-ethanolamine, eicosapentaenoyl-ethanolamine, docosahexaenoyl-ethanolamine, or lipids of all three omega-3 acids).
  • DHA docosahexaenoic acid
  • the DHA ends up in the neuronal membrane in form of phospholipids. If the DHA is synthesized in the liver or administered intravenously, then it apparently crosses the BBB to the brain.
  • the DHA enrichment in the brain may result from three important roles of the BBB: (1) to take up ALA from the blood, transform it into EPA and eliminate the latter into the perivascular glial cells of the brain, (2) take up DHA from the blood and release it into the brain and (3) to block the egress of DHA from the brain.
  • the latter is probably responsible for the relatively high DHA content of brain phosphotriglycerides when compared to other polyunsaturated fatty acids (N. Salem et al., in Health Effects of Polyunsaturated Fatty Acids in Seafoods. A.P. Simopoulos, ed. Academic Press, N.Y., 1986, p. 263-317 and G.Y. Sun and L.L.
  • 4,935,465 teaches the attachment of one or more polymer molecules to a protein.
  • attachment of polymers to proteins particularly to enzymes which act on low molecular weight substrates, retards enzyme clearance and decreases enzyme antigenicity.
  • the enzyme acts on a macromolecular substrate or on a cell-bound substrate, the enzyme activity of the conjugate could be diminished.
  • U.S. Pat. No. 4,046,722 discloses anti-cancer drugs covalently bonded to cationic polymers for the purpose of directing them to cells bearing specific antigens.
  • the polymeric carriers have molecular weights of about 5,000 to 500,000.
  • Further work involving covalent bonding of an agent to a cationic polymer through an acid-sensitive intermediate (called also spacer) molecule is described in U.S. Pat. No. 4,631,190 and U.S. Pat. No. 5,144,011.
  • spacer molecules such as cis-aconitic acid, are covalently linked to the agent and to the polymeric carrier.
  • the drug can be selectively hydrolyzed from the molecular conjugate and released in the cell in its unmodified and active form.
  • Molecular conjugates are transported to lysosomes, where they are metabolized under the action of lysosomal enzymes at a substantially more acidic pH than other compartments or fluids within a cell or body.
  • the pH of a lysosome is shown to be about 4.8, while during the initial stage of the conjugate digestion is possibly 3.8.
  • U.S. Pat. No. 5,308,701 discloses a method for encapsulating proteins within a synthetic cationic poly-L-lysine, crosslinked with multivalent ions of the opposite charge, to form a hydrolytically stable gel.
  • Synthetic cationic polymers are more suitable drug carriers than natural polymers because their structure can be systematically altered in a defined way and therefore, it is possible to design them to suit biological requirements such as penetration through the blood-brain barrier.
  • the polymeric cationic carrier system used to facilitate the crossing of the blood-brain barrier comprises poly-L-lysine (PLL).
  • PLL is a bio- compatible, hydrophilic polymer with very thoroughly studied chemical and biological properties.
  • Other cationic polyamino acids such as polyarginine and polyornithine are within the scope of this invention.
  • These polyamino acids are covalently attached to a biopolymeric agent conjugated with omega-3 fatty acid molecules or other polymeric protecting groups (e.g., polyethylene glycol).
  • omega-3 fatty acid molecules or other polymeric protecting groups e.g., polyethylene glycol
  • a nontoxic fragment of a neurotoxin is further attached to assure targeting and internalization by neurons once the neuronal target is reached. Spacers can be inserted between the components of the vehicle as shown in the description of the preferred embodiments.
  • one or more fatty acid moieties can be attached to the enzyme by reacting an imidate activated enzyme with the respective fatty acid.
  • the number of attached moieties is limited only by the number of available -NH 2 groups of the enzyme.
  • the prior art is silent on lipophilic macromolecular conjugates, potentially containing polycationic amino acids (targeting lipophilic macromolecular vehicles), used to deliver therapeutic or diagnostic agents across the BBB selectively to specific areas of the brain.
  • a major object of the present invention is to synthesize site-specific biomolecular complexes for the selective transport of a therapeutic, prophylactic and diagnostic agent to the target brain cells.
  • the lipophilic components of the macromolecular complex or conjugate are biocompatible with tissue components.
  • the targeting complexes are further specific for agent delivery through receptor pathways to neuronal membranes or to astroglial cells.
  • Preferred features of the present invention are the synthesis and use of lipophilic targeting complexes to deliver the "required agent dose" in vivo to specific areas of the brain, in controlled therapeutic levels by intravenous infusion, intrathecal injection into the cerebrospinal fluid (CSF), or direct infusion into the brain interstitial fluid (BIF).
  • CSF cerebrospinal fluid
  • BIF brain interstitial fluid
  • the instant invention describes a series of site-specific therapeutic, prophylactic or diagnostic complexes comprising a biologically active molecule (agent), one or more residues of an omega-3 fatty acid or derivatives thereof, a polyamino acid as a cationic carrier and a neuron-targeting, internalizing moiety.
  • a biologically active molecule agent
  • residues of an omega-3 fatty acid or derivatives thereof a polyamino acid as a cationic carrier
  • a neuron-targeting, internalizing moiety By complex or conjugate is meant a molecule that is either held together by van der Waal's and electrostatic interactions or by covalent bonds.
  • the preferred omega-3 fatty acids are eicosapentaenoic acid (EPA), docosahexaenoic acid (DHA) and alpha- linolenic acid (ALA).
  • the specific bonding in the complex will be dictated by the specific chemistries used to form the complex of the invention.
  • the therapeutic complexes of the present invention are particularly useful for treating diseases that occur in the glial tissue of the brain and diseases that occur in the cortical, cholinergic and adrenergic neurons of the central nervous system.
  • the biologically active agent is selected from the group consisting of amino acids, peptides, poly peptides, proteins, fusion proteins, polyethylene glycol- derivatized proteins, enzymes, hormones, growth factors, antigens, antibodies, nucleic acids and derivatives thereof, DNA- and RNA-segments, oligonucleotides, antisense oligonucleotide analogs, nucleotides and derivatives thereof, genes, transfected cells, biological vectors, cloning vectors, expression vectors, neurotoxins and fragments thereof, and diagnostic agents.
  • the biological agent can further be derivatized with peptide residues called permeabilizer peptides for facilitated crossing of the blood- brain barrier.
  • the site-specific complex of the present invention may optionally further comprise a polyamino acid such as polylysine, polyarginine, polyornithine and polyasparagine.
  • a polyamino acid such as polylysine, polyarginine, polyornithine and polyasparagine.
  • the preferred polyamino acid is a polylysine having 1-50 lysine residues and preferably from 10 to 25 lysine residues.
  • These polyamino acids can further be derivatized with omega-3 polyunsaturated fatty acids (ALA, EPA or DHA) and permeabilizer peptides for facilitated crossing of the blood-brain barrier.
  • omega-3 polyunsaturated fatty acids ALA, EPA or DHA
  • permeabilizer peptides for facilitated crossing of the blood-brain barrier.
  • the site-specific complex of the present invention may optionally further incorporate a neuronal targeting and/or internalizing moiety selected from the group consisting of the C-fragment of tetanus toxin (TTC), of nontoxic fragment of alpha- bungarotoxin (ABT), other neuron-specific toxins and their nontoxic fragments that still retain their directing and internalizing capacity, and nerve growth factor (NGF).
  • TTC C-fragment of tetanus toxin
  • ABT nontoxic fragment of alpha- bungarotoxin
  • NGF nerve growth factor
  • the instant invention is further inclusive of methods for the site-directed delivery of drugs to treat diseases of the glial tissue of a mammalian brain by intrathecally administering to the afflicted mammal a therapeutically effective amount of a biologically active agent, called also drug, that has been derivatized with eicosapentaenoic acid.
  • a biologically active agent called also drug
  • the active agents are derivatized with docosahexaenoic acid.
  • the instant invention is further inclusive of methods for the site-directed delivery of the above complex/conjugate by direct infusion into the interstitial matter of the brain of the afflicted mammal.
  • the diagnostic agent is derivatized accordingly, i.e., eicosapentaenoic acid when glial delivery is required and docosahexaenoic acid when cortical, cholinergic and adrenergic neuron delivery is required.
  • a targeting/internalization moiety might also be included in the complex through direct attachment to the agent.
  • the present invention further describes a transport vector comprising the above site-specific moiety covalently bonded to a cationic poly-L-lysine carrier having a molecular weight of 1,000 to 50,000 and further bonded (directly or indirectly) to one or more polyunsaturated fatty acid residues.
  • the cationic polyamino acid carrier moiety can be bonded to a phospholipid or diglyceride.
  • the transport vector which contains the site-specific complex and the carrier can contain 1 to 5 molecules of an active agent, (such as an enzyme), or 1 to 5 site-specific complex units (molecules), by random coupling to pendant epsilon-amino groups of the carrier. Other coupling methods are also within the scope of the invention.
  • the molecular weight of the poly-L-lysine carrier By controlling the molecular weight of the poly-L-lysine carrier, it is possible to regulate the penetration of the whole macromolecular vehicle (cationic at physiological pH) through the BBB in vivo, and to predict, design and fabricate the adequate drug delivery system with high performance in vivo.
  • site-specific complexes contain cationic carriers, intravenous administration into mammals is preferred.
  • the site-specific complex of the present invention which includes the active agent and cationic carrier will further contain one or more DHA or EPA moieties so that the complexes may reach their sites of action more effectively.
  • the number of DHA or EPA moieties in such complexes may vary from 1 to 20 moieties.
  • the instant invention is further inclusive of methods for the site directed delivery by means of the above described transport vector of drugs to treat diseases of the glial tissue of a mammalian brain by intravenous administration and penetration through the blood-brain barrier (of an afflicted mammal) of a therapeutically effective amount of a biologically active agent also called drug, that has been derivatized with alpha-linolenic acid.
  • a biologically active agent also called drug
  • the omega-3 fatty acid used for derivatization is DHA.
  • three different groups are attached to the main polymeric carrier-chain comprising a biodegradable homopolymer of poly-L-lysine.
  • One or more molecules of the active agent (which is the entity that initiates the therapeutic or physiological response). These become active only after being cleaved from the vehicle.
  • the attachment of the agent can be to the amino terminal of the main chain of the poly-L-lysine carrier or through epsilon-amino groups of L- lysine components of the polymeric carrier (e.g., a side spacer) to the main polymer chain.
  • the agent is an enzyme or a protein, in a protected form, with low antigenicity following administration to mammals.
  • a targeting and/or internalizing moiety can be attached to poly-L-lysine through an epsilon-amino group of L-lysine.
  • This moiety is a specific transport moiety to neuronal brain tissue or cells.
  • targeting moieties are pH- sensitive groups or receptor-active components, such as antibodies or neurotoxin fragments compatible with a receptor on a specific cell surface, (e.g.
  • TTC tetanus toxin fragment-C
  • ABT nontoxic alpha- bungarotoxin
  • NTF nerve growth factor
  • TTC tetanus toxin fragment-C
  • ABT nontoxic alpha- bungarotoxin
  • NEF nerve growth factor
  • One or more hydrophobic groups comprising omega-3 fatty acids or their derivatives can also be attached to the macromolecular carrier by covalent bonds to enhance the lipophilic character of the polar agent transport vehicle. Increased lipophilicity aids incorporation of the complex into the lipid matrix of the microvascular endothelial membrane constituting the blood brain barrier.
  • the instant invention further comprises site-directed complexes, wherein the lipophilic moiety derived from omega-3 fatty acids or their derivatives is attached to the biologically active agent, such as enzymes, through electrostatic forces.
  • site-specific complexes wherein said complex molecules are held together through van der Waals forces are within the scope of this invention.
  • Such complexes might have the advantage over the covalently bound conjugates of facilitating the bioavailability of the agent transported into a specific area of the brain without requiring a chemical reaction to achieve the release of the agent from the conjugate.
  • the resultant site-specific complex interacts with water to form self-assembling structures in colloidal dispersions.
  • the site-specific complex is mixed with lipids, such as phosphatidyl choline and cholesterol, in aqueous systems, to form liposomes by conventional methods.
  • lipids such as phosphatidyl choline and cholesterol
  • the pharmaceutical preparations are administered in vivo where the said complex is attracted to the specific cell membrane and its uptake to the brain is enhanced. Therefore, the colloidal particles (e.g., micelles, vesicles or liposomes) are specifically targeting a cell surface determinant and subsequently internalized through endocytosis or pinocytosis.
  • this invention comprises a method for facilitating the transport of an agent into brain cells with said stable colloidal aqueous dispersion of site-specific complexes of the agent.
  • the lipophilic group of the site-specific complex is a hydrophobic moiety which allows insertion of the complex into the cell membrane or into liposomal bilayers.
  • the cell surface-bound complex is internalized, and the agent is released into the cell.
  • Enzymes and proteins which are defective in lysosomal storage diseases are listed in Table 2, (entitled “Lysosomal Storage Diseases").
  • the site specific complexes of listed active enzymes and proteins are intended to be utilized in the therapy of diseases mentioned in Table 2, and they are within the scope of the invention.
  • the enzyme is human glucocerebrosidase (or its recombinant form), or modifications thereof, such as alglucerase or ceredase and cerezyme.
  • a fusion protein e.g., viral fusion proteins, influenza hemagglutinin
  • covalently coupled with the lipophilic moiety through the polylysine carrier or as a targeting moiety of the agent complex in delivery system enhances endocytosis of colloidal particles which are ultimately broken down in the lysosomes.
  • the fusion proteins e.g., viral envelope glycoprotein of Sendai virus, influenza virus or herpes Simplex virus
  • any of the included complexes of this invention can be administered intrathecally, directly infused into the brain or administered intravenously.
  • FIG. 1 shows a model of a lipophilic polylysine as an amphipathic conjugate or complex;
  • DAPS diacyl phosphatidyl serine
  • lyso-phosphatidyl serine may be used.
  • phosphatidyl ethanol amine may be used.
  • R j and R 2 represent alkyl chains of fatty acids of omega-3 or omega-6 series.
  • FIG. 2 is a schematic representation of a lipophilic site-specific biomolecular complex or a targeting vehicle for pharmacologic agents designed by covalent linkage of a fatty acid or a phospholipid molecule (noted as lipophilic moiety: LP) to the agent (A) through a poly-L-lysine (PLL, wherein subscript N has values between zero and 50, preferably 11 lysyls).
  • said complex is further coupled with another segment of poly-L-lysine (PLL, wherein subscript M comprises values between zero and 50, preferably 11 lysyls) derivatized with a targeting moiety (T) for selective agent delivery to the brain.
  • FIG. 3 is a schematic representation of a site-specific biomolecular complex or a targeting vehicle for pharmacologic agents constructed by covalent linkage of a fatty acid or phospholipid molecule (represented by lipophilic moiety) to the agent (A) through a poly-L-lysine (PPL, wherein subscript N is between zero and 11), optionally coupled with another segment of poly-L-lysine (PPL, wherein subscript M is an integer having values between zero and 11) derivatized with a targeting moiety for selective drug delivery to the brain.
  • PPL poly-L-lysine
  • subscript M is an integer having values between zero and 11
  • Phosphatidyl serine is coupled to main polymeric chain through its -COOH group.
  • Phosphatidyl serine is coupled to main polymeric chain through its -NH 2 group.
  • Subscript N has values between 1 and 25.
  • FIG. 7 shows a site-specific biomolecular complex or a targeting vehicle of a biologically active agent, wherein the agent is derivatized with a lipophilic carrier.
  • Targeting moiety is shown in the same fitrure (b).
  • the lipophilic functional poly-unsaturated molecules which form part of the site-specific complex are preferably selected from the group consisting of fatty acids of the omega-3 series or lipid derivatives thereof.
  • Other examples of lipophilic molecules are fatty acids, diacyl glycerols, diacyl phospholipids, lyso-phospholipids, cholesterol and other steroids, bearing poly-unsaturated hydrocarbon groups of 18 to 46 carbon atoms.
  • Preferred bio-polymer carriers are poly (alpha)-amino acids (e.g., PLL, poly L-arginine:PLA, poly L-ornithine:PLO), human serum albumin, aminodextran, casein, etc. These carriers are biodegradable, biocompatible and potentially excellent candidates for drug delivery systems.
  • the macromolecular carrier is a polypeptide carrier exemplified by poly-L- lysine which is represented by the following structural formula:
  • poly-lysine carrier contains the lysines, either D- or L-configuration, linked alternatively by typical peptide bonds to each other.
  • polypeptide that is within the scope of this invention is a C-terminal and N-terminal arginine or the masked or blocked arginine.
  • Preferred N-terminal amino acid groups are arginine, acetyl arginine, lysine or acyl lysine, where these terminal amino acids are of either D- or L-configuration.
  • the amino acids that constitute the core sequence of the polypeptide of this invention are preferably in the L-isomer form. Characteristic features and structural/colloidal behavior of the polypeptides of this invention are essential to allow the proper polymer configuration/conformation to effect an increase of permeability of BBB when they are attached to a drug administered to the host animal.
  • the host can be humans, domestic animals (e.g., dog, cat, horse, etc.), and animals for experimental purposes (e.g. , rabbits, rats and mice) or any animal which possesses a central nervous system (i.e., a brain).
  • animals for experimental purposes e.g. , rabbits, rats and mice
  • a central nervous system i.e., a brain
  • the D-isomer of lysine is optionally added for one or more times in the linear core sequence of lysines, particularly in C-terminal region, the penetration of the drug-conjugates through the BBB might be altered, even diminished.
  • the polylysine should be derived from the L-isomer.
  • Known preparative methods can be used for synthesis of PLL, with desired average molecular weight: MW.
  • hydrobromide salts of PLL of varying MW's are also commercially available, for example, hydrobromide salts of PLL of MW 3300-4000 (i.e., PLL with about 26 lysyls chain length) and of MW 180,000 - 260,000 (i.e. , 1200-1700 lysyls in poly-lysine chain length), etc.
  • polypeptides with less than 20 amino acids are prepared by known solid phase synthesis.
  • a preferred embodiment of this invention is poly-L-lysine, noted also as
  • PLL or poly-Lys with n comprised between 6 and 25, sometimes at least 50 lysine molecules as a constituent part of a site-specific complex.
  • an artificial model system i.e. , a drug vehicle
  • lipophilic poly-Lys carrier for drug delivery systems.
  • This invention also pertains to lipophilic poly-Lys conjugates or complexes wherein a wide variety of lipophilic compounds (e.g., highly unsaturated fatty acids or derivatives thereof) are covalently bound to linear ( Figure 1) or branched poly-Lys.
  • lipophilic compounds e.g., highly unsaturated fatty acids or derivatives thereof
  • Figure 1 linear or branched poly-Lys.
  • highly unsaturated fatty acid derivatives such as phospholipids are, but not limited to those, listed in Table 1.
  • the acyl derivatives of phosphatidyl serine (DAPS) are not critical, but in general (see, Table 1) will contain at least 16 carbon atoms.
  • the fatty acyls Ri and R 2 contain from 16 to 24 carbon atoms exclusive of additional side chains or functional groups.
  • the fatty acyls may be saturated (in snl-position of glycerol backbone) and poly-unsaturated of omega-3 or omega-6 essential fatty acid series (in sn2-position) or both poly-unsaturated with at least 18 carbon atoms.
  • Suitable fatty acids include palmitic (PA), stearic (STA), linoleic (LO), linolenic (LA), arachidonic (AA), di-homo gamma linolenic (DGLA), eicosapentaenoic (EPA) and docosahexaenoic (DHA) acids.
  • Other lipophilic poly-lysine conjugates can be formed through one or more peptidic bonds between amino group(s) of the carrier and carboxylic group(s) of a poly-unsaturated fatty acid(s) of omega-3 or omega-6 series, preferably DHA, EPA and DGLA.
  • lipophilic poly-Lys is by using branched poly- Lys, for example with a single lysine like a free block.
  • the lipophilic-PLL carrier of pharmaceutical compositions of this invention can also contain neuropharmaceutical agents which have a prophylactic and/or therapeutic effect on neurological disorders.
  • neurological disorders include: malignant brain tumors, autoimmune deficiency syndrome (AIDS), Parkinson's disease, neurodegenerative disease, Alzheimer's disease, multiple sclerosis, migraine, pain or a seizure disorder, epilepsy, depression and trauma, neuronal storage disease, and other severe deterioration of the CNS.
  • This invention also relates to the design of novel drug-vehicles or drug-shuttles with increased lipophilicity and increased penetration into biological membranes having facilitated transport throughout the body.
  • the invention is also particularly useful for delivering to neuronal sites pharmaceutically active agents such as anti- neoplastic agents, anti-microbial agents, anti-parasitic agents, adrenergic agents and catecholaminergic agents, anti-convulsants, nucleotide analogues, anti-trauma agents, enzymes and proteins used to prevent or treat neurological disorders.
  • pharmaceutically active agents such as anti- neoplastic agents, anti-microbial agents, anti-parasitic agents, adrenergic agents and catecholaminergic agents, anti-convulsants, nucleotide analogues, anti-trauma agents, enzymes and proteins used to prevent or treat neurological disorders.
  • targeting lipophilic complexes (presented in Figure 2), both the therapeutic agent and targeting moiety are attached at the terminals of the main linear
  • PUFA is chosen from a group consisting of omega-3 essential PUFA, more preferably alpha-linolenic acid (ALA), eicosapentaenoic acid (EPA) and most preferably docosahexaenoic acid (DHA) and represented by the formula Rt-COOH.
  • omega-3 essential PUFA more preferably alpha-linolenic acid (ALA), eicosapentaenoic acid (EPA) and most preferably docosahexaenoic acid (DHA) and represented by the formula Rt-COOH.
  • the enzyme derivatives of the present invention are particularly useful for treating lysosomal storage diseases which cause severe neurodegeneration and are manifested by enzyme deficiencies.
  • the specific enzymes which can be derivatized are, but not limited to those, listed in Table 2 [Neufeld, Annu. Rev. Biochem., 60:257-80 (1991)].
  • r is an integer from 1 to 20 which shows the varying numbers of lipophilic residues attached to the enzyme macromolecule and is controlled primarily by the ratio of PUFA to lysyls of enzyme chain length and secondarily by the duration of the coupling reaction.
  • the composition and structure may be determined for any condensation product by the known method in the art (Lowry et al. , J. Biol. Chem., 193, 265-275 (1951)).
  • the conditions for reaction [2] are conventional conditions for the formation of an amide linkage, such as by activating the fatty acid (HOOC-R j ) with a suitable activator under non-aqueous conditions, for example dicyclohexylcarbodiimide, and subsequent reaction with the appropriate amine group(s) on enzyme.
  • a suitable activator under non-aqueous conditions, for example dicyclohexylcarbodiimide, and subsequent reaction with the appropriate amine group(s) on enzyme.
  • pH may be controlled, if necessary, by acid or base addition either manually or automatically.
  • an enzyme is dissolved in an aqueous buffer preferably at a moderately alkaline pH 7-9.5 (e.g., phosphate buffered saline: PBS).
  • a moderately alkaline pH 7-9.5 e.g., phosphate buffered saline: PBS.
  • the solution of enzyme is treated with 0.6 equivalents of PUFA dissolved in non-aqueous medium in the presence of 1 equivalent of suitable carbodiimide, preferably dicyclohexylcarbodiimide, under the argon and suitable anti-oxidant compounds to protect from damage the PUFA during the conjugation reaction.
  • the reaction is permitted to proceed for about 17 hours at ambient temperature. Then the reaction mixture is passed through Sephadex columns.
  • the reaction product provides around 2 PUFA residues per enzyme, (see formula
  • Carbodiimide coupling is a well-known procedure outlined in the following literature references: Sheehan, J. C. and Hess, G.P. , "New method of forming peptide bonds", J. Am. Chem. Soc, 77, 1067 (1955); Halloran, M.J. and Parker, C. W. , "The preparation of nucleotide-protein conjugates: Carbodiimides as coupling agents", J. Immunology, 96, 373 (1966); Kurzer, F., and DouraghiZadeh, K.,
  • Method for preparation of enzyme covalently coupled with PUFA using the conjugation reaction between enzyme and N-hydroxysuccinimide ester of PUFA in phosphate buffered saline (PBS) at pH 6-9.5, containing deoxycholate For example, 3 mg of purified enzyme is added to 50 microgrammes of N-hydroxysuccinimide ester of docosahexaenoic acid in PBS containing 2-3% deoxycholate. The mixture is incubated at 37 °C for 9 to 15 hrs and then chromatographed on a Sephadex G-75 column (1.3 x 45 cm) in PBS containing 0.15 % deoxycholate to remove excess of PUFA.
  • PBS phosphate buffered saline
  • Conjugation of the enzyme with fatty acids of omega-3 series can be carried out according to known procedures for reacting amino groups on enzyme with carboxyl group of poly-unsaturated fatty acids (as shown in example 1) or with their acyl chlorides or anhydrides.
  • enzyme is dissolved in bicarbonate buffer (BB, of pH 9); then PUFA-anhydride, dissolved in anhydrous dimethyl sulfoxide (DMSO), is added immediately to the enzyme solution.
  • Enzyme- PUFA complexes of formula [1] are prepared. The stoichiometric ratio of PUFA/Enzyme could be varied as justified already.
  • the reaction is allowed to proceed for 2 hours at 25 °C. Then, the reactants were passed through G-25
  • reaction product of an enzyme derivatized with PUFA and chemically linked to a protective moiety to decrease the enzyme antigenicity, e.g., human serum albumin (SA) or polyethylene glycol (PEG), is a non-immunogenic, protected-enzyme-PUFA conjugate.
  • SA human serum albumin
  • PEG polyethylene glycol
  • T is a targeted non-immunogenic, protected enzyme-PUFA vehicle, wherein T stands for the targeting moiety for specific delivery of the resultant conjugates to selective cellular surface receptors.
  • T is selected from the group consisting of tetanus toxin fragment C (TTC), nerve growth factor (NGF), alpha-bungarotoxin (ABT), cell- surface directed antibodies (such as immunoglobulin G, antibody against human pancreatic tumor cells, and antibody against hepatocytes) and insulin as an example of a serum hormone used as targeting moiety, depending on the desired site of delivery.
  • TTC tetanus toxin fragment C
  • NVF nerve growth factor
  • ABT alpha-bungarotoxin
  • cell- surface directed antibodies such as immunoglobulin G, antibody against human pancreatic tumor cells, and antibody against hepatocytes
  • insulin as an example of a serum hormone used as targeting moiety, depending on the desired site of delivery.
  • T-S-SA-S-E-(PUFA) r li2> ...,20 r ⁇
  • the resultant molecular conjugate [6] is designed for specific cytoplasmic delivery of the enzyme.
  • T tetanus toxin fragment C :TTC or poly L- lysine.
  • PLL PLL are used as examples, but not limited to them. PLL could be used as a targeting moiety in terms of effecting cellular uptake of its molecular conjugates to an agent (enzyme) and also as a cationic transport carrier under the physiological conditions.
  • T-E-PUFA conjugates [7] are synthesized by carbodiimide coupling procedure. Also, the T-E-PUFA conjugates can be achieved by a complex method, using glutaraldehyde coupling procedure and carbodiimide coupling procedure, and the resulting conjugates are given by [8] :
  • the E-T-PUFA conjugates are synthesized by carbodiimide coupling procedure. Also, the E-T-PUFA conjugates [10]:
  • the E-PLL-PUFA conjugates represented by [11] are achieved by carbodiimide coupling procedure. Also, similar E-PLL-PUFA conjugates given by - [12]:
  • T-E-PLL-PUFA molecular conjugates are prepared in three steps using carbodiimide coupling. Also, a mixed method using a combination of glutaraldehyde coupling procedure and carbodiimide coupling procedure could be achieved and the resulting T-E-PLL-PUFA molecular conjugates are illustrated by [14]:
  • the resulting conjugates of [15] could be symbolized by E-PLL(T)-PUFA.
  • a pH-sensitive spacer molecule (noted as S) can be inserted between agent (such an enzyme) and PLL carrier or between other parts of the transport vehicles, depending on the desired properties of the resulting final conjugate. Examples are illustrated by [16] and [17]:
  • conjugation can be achieved by chemical reactions, using step reaction procedures, well known in the art, employing carbodiimide catalyst, or combinations of glutaraldehyde coupling and carbodiimide coupling modes.
  • PUFA i.e., E-(NH-OC-R ⁇ ) r , as shown in examples 1 to 3.
  • step II the conjugated product of step I, E-(NH-OC-R j ) r , is covalently coupled with a specific target moiety.
  • This reaction is carried out according to known procedures for reacting amino groups on enzyme with carboxyl groups of the targeting moiety to provide linkage between them, for example using carbodiimide coupling as in step I (see, example 1).
  • Other modes of conjugation can be used, depending on the structure and properties of the individual molecules employed to build the vehicle.
  • bifunctional protein modifying reagents such as glutaraldehyde
  • the conjugated T-E-PUFA product can also be achieved by the following reactions:
  • the conjugation reaction [18] may be achieved using a variety of bifunctional protein modifying reagents.
  • reagents include: N-succinimidyl-3-(2- pyridyldithio)propionate, active esters such as disuccinimidyl suberate, aldehydes such as glutaraldehyde used in reaction [18].
  • Glutaraldehyde coupling is described in the prior art: Avrameas, S. and
  • reaction [18] is further derivatized with PUFA, following the procedure of reaction [19].
  • conjugates of formulae [9] to [17] can be made in the intermediate steps, depending on the enzyme placement in the conjugated individual vehicle molecule.
  • targeting moiety tetanus toxin fragment C :TTC can be used as an example, but not limited to.
  • PUFA such as DHA
  • PUFA/PLL a chosen stoichiometric ratio PUFA/PLL of two.
  • the molecular complex of (DHA) 2 -PLL is purified as known in the art.
  • phosphatidyl serine (rac)-l- O-octadecanoyl, 2-O-docosahexaenoyl-glycero-3-phospho-serine, named also l-O- stearoyl, 2-O-docosahexaenoyl-glycero-3-phospho-serine:SDPS) is substituted for the
  • the reactants of example 1 are used, except that molecular conjugate of poly ⁇ lysine with DHA (product of example 4) is used to substitute for the PUFA.
  • the quantity of said reactants is calculated for a chosen stoichiometric ratio (see formula [9]) within the desired limits for the hydrophilic hydrophobic balance of the resultant transport vector of enzyme.
  • the final molecular complex of lipophilic enzyme must be present with at least 70% of cationic charges initially available (unmodified PLL) for excellent uptake into cell.
  • PLL-(SDPS) 2 conjugates or complexes are synthesized by coupling SDPS to the alpha-amino group and to the carboxyl group of a suitably protected epsilon-amino
  • PLL molecule e.g., about 12 lysyls in a PLL chain length
  • carbodiimide coupling procedure in a similar manner as in example 4.
  • the reactants of example 4 are used, except that sodium salt of phosphatidyl serine ((rac)- 1 -O-octadecanoyl, 2-O-docosahexaenoyl-glycero-3-phospho-serine, named also 1-O-stearoyl, 2-O-docosahexaenoyl-glycero-3-phospho-serine:SDPS) is substituted for the PUFA with the stoichiometric ratios of 2 for SDPS/PLL-macromolecule and
  • PLL molecule has epsilon-amino groups protected.
  • the coupling reaction entailed adding a 2% solution of SDPS in ethanol in aliquots to a stirred 0.1 % solution of the modified (epsilon-amino protected) PLL in 0.01 mol/1 carbonate buffer at pH 9, or 0.01 mol/1 phosphate buffer at pH 7.5 in the carbodiimide presence, at 0°C.
  • SDPS to PLL is two (i.e. , 1 SDPS to 6 lysyls), and of buffer to ethanol, 100: 1 (v/v).
  • the molecular conjugate of the first step, enzyme derivatized with PUFA, such as DHA, described in examples 1-3 is covalently coupled, in a second step of conjugation reaction, to a specific targeting moiety, such as tetanus toxin fragment C (TTC).
  • TTC tetanus toxin fragment C
  • the second conjugation is carried out according to known procedures, for example, reacting amino groups on enzyme with carboxyl groups of targeting moiety (if available) to provide linkage between them.
  • the molecular conjugate of the first step, PLL derivatized with PUFA, such as DHA, described in example 4, is covalently coupled, in a second step of conjugation reaction, to a specific enzyme.
  • the second conjugation is carried out according to known procedures, for example, reacting amino groups on PLL with carboxyl groups of enzyme (if available) to provide linkage between them.
  • crystalline enzyme glucocerebrosidase
  • poly-L-lysine of molecular weight of 6700
  • EDC l-ethyl-3-(3- dimethyl-aminopropyl)carbodiimide
  • This solution was incubated at room temperature (around 22 °C) for 7 hours under argon with occasional shaking, and then loaded onto a Sephadex G- 100 chromatographic column which had been previously equilibrated with 0.01 M phosphate buffered saline (PBS), pH 7. After loading, the column was eluted with PBS and fractions containing enzyme coming out of the column at and around the void volume were collected, pooled, concentrated to a volume of 1.0 ml, and then diluted with water to a volume of 10 ml. In order to remove unreacted enzyme, this solution was passed through a DEAE-Sephadex column.
  • PBS phosphate buffered saline
  • HRP horseradish peroxidase
  • the HRP-PLL-DHA conjugates were used in experiments.
  • the enzymatic activity of the pooled HRP-PLL-DHA conjugates was compared to that of unconjugated HRP using an assay employing dianizidine as an electron acceptor (see, U.S. Pat. No. 4,847,240). It was found that the conjugation decreased the enzymatic activity of HRP by about 40%.
  • E-NH-OC-CH C(COOH)-CH 2 -CO-NH-PLL-(NH-OC-R 1 ), [20]
  • E-NH-OC-CH C(COOH)-CH 2 -CO-NH-PLL-(NH-OC-R,) [21]
  • spacer molecules which contain more than one maleic anhydride ring, i.e., poly maleic anhydride, also commercially available.
  • the molecular conjugate of example 12 presents the advantage that is pH sensitive and in mild acidic conditions will release easily the enzyme in unmodified form, which is in a biologically active state. Using these molecular conjugate of example 12 it is possible to control the intracellular release of the enzyme (e.g. , within the lysosome).
  • each pair of cis-carboxyl groups formed by hydrolyzing a maleic anhydride is believed to comprise a suitable location for bonding an enzyme molecule. Therefore, numerous enzyme molecules may be bonded to a single polymeric spacer molecule, as shown in formula [22]:
  • the enzyme selected to further exemplify the present invention is the native acid beta-glucosidase (ABG) or glucocerebrosidase, its mannose-terminated derivative alglucerase (mABG) or ceredase, its recombinant acid beta-glucosidase from (rABG) and its recombinant manose-te ⁇ ninated form cerezyme (mrABG). It appears to be five reactive epsilon amine groups of lysines in each form of the enzyme, i.e., in
  • ABG, mABG, rABG, and mrABG can be partially or fully docosahexaenoylated (DHA) or eicosapentaenoylated (EPA).
  • DHA docosahexaenoylated
  • EPA eicosapentaenoylated
  • ABG -(CO-S-DHA) ⁇ _ 5 wherein S is a diamine like spacer ethylene diamine (NH 2 -CH 2 -CH 2 -NH 2 ) which is the shortest spacer, and
  • the five epsilon amine groups of the enzyme's lysine residues can also be derivatized with DHA-containing phosphatidyl serine, through the serine carboxyl group and the formation of an amide bond (see Table 1).
  • ABG exemplifies all known forms of the enzyme: ABG, rABG, mABG and mrABG.
  • ABG*-(NH-DPPS),_ 5 and ABG* (NH-DDPS), ⁇ wherein DPPS is 2-docosahexaenoyl, 1-palmitoyl-phosphatidylserine, and DDPS is 1 ,2-didocosahexaenoyl-phosphatidylserine. Further examples are given with aspartic or glutamic acid residues derivatized with DHA-containing phosphatidyl-ethanolamine:
  • DPPE is 2-docosahexaenoyl, 1-palmitoyl-phosphatidylethanolamine and DDPE is l,2didocosahexaenoyl-phosphatidylethanolamine.
  • the DHA-containing phospholipids can also form complexes with the various forms of acid beta-glucosidase. Examples are represented below: [ABG * ],_ 5 [DPPS], .5 , [ABG * ],. 5 [DDPS] ⁇ 5 ,
  • an epsilon amine group of the enzyme's lysine residue is attached to the carboxyl terminal of the polycationic carrier (polylysine chain: (Lys) ⁇ . 30 , and DHA or DHA-containing phospholipid to the amine terminal of the polylysine chain.
  • a directing moiety such as tetanus toxin fragment C(TTC) or nerve growth factor (NGF) attached to an apsilon amine of the polylysine carrier provide another group of site-specific biomolecular complexes or shuttle vectors with therapeutic uses.
  • the direction moiety is coupled to the docosahexaenoylated enzyme directly or through a diamine spacer.
  • the directing moieties TTC and NGF can themselves be docosahexaenoylated.
  • TTC is directive to neurons of the central nervous system and it is internalized by cortical neurons of the gray matter. Thus, it provides the shuttle vector with the additional capability of neuronal endocytosis.
  • NGF distributes throughout the central nervous system, but is appears to concentrate in the vicinity of neurons of the cholinergic nervous system including neurons responsible for cogmtive and memory processes such as the basal ganglia, the nucleus of Meynart, and the hippocampus in general. Further examples are:
  • TTC-(NH DHA)p NGF-(NH-DHA) r p is the number of epsilon amine groups of lysines in TTC derivatized with DHA and r. The number of epsilon amine groups of lysines in NGF derivatized with DHA.
  • Examples 1 to 13 can be extended by using a wide variety of phospholipids, such as phosphatidyl ethanol amine, acylated phospholipids, lysophospholipids and or diacyl glycero derivatives and phosphatidic acids with the proviso that PUFA chains are in sn2 position or in both snl and sn2 position, if they are available.
  • phospholipids such as phosphatidyl ethanol amine, acylated phospholipids, lysophospholipids and or diacyl glycero derivatives and phosphatidic acids with the proviso that PUFA chains are in sn2 position or in both snl and sn2 position, if they are available.
  • agent such as enzyme, proteins, factors, cofactors, hormones, anti-cancer drugs, anti- Parkinson's drugs, etc.
  • carrier such as PLL of varying molecular weights, different lipophilic residues, and optionally targeting moieties

Landscapes

  • Health & Medical Sciences (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Engineering & Computer Science (AREA)
  • Bioinformatics & Cheminformatics (AREA)
  • Proteomics, Peptides & Aminoacids (AREA)
  • Public Health (AREA)
  • Chemical & Material Sciences (AREA)
  • Medicinal Chemistry (AREA)
  • Pharmacology & Pharmacy (AREA)
  • Epidemiology (AREA)
  • Animal Behavior & Ethology (AREA)
  • General Health & Medical Sciences (AREA)
  • Veterinary Medicine (AREA)
  • Molecular Biology (AREA)
  • Toxicology (AREA)
  • Medicines That Contain Protein Lipid Enzymes And Other Medicines (AREA)
  • Medicinal Preparation (AREA)

Abstract

La présente invention se rapporte à des complexes biomoléculaires dirigés comprenant un agent thérapeutique, prophylactique et diagnostique, appelé également molécule biologiquement active, et à un acide gras oméga 3 et ses dérivés. Les complexes sont également liés de manière covalente à des porteurs cationiques et à des peptides perméabilisant de sorte qu'ils puissent être administrés dans la barrière hémato-encéphalique, ainsi qu'à des fractions de ciblage destinées à être capturées par des cellules neuronales cibles. Ces complexes sont notamment utilisés pour administrer un agent biologiquement actif dans le tissu glial du cerveau ainsi que dans les neurones corticaux, cholinergiques et adrénergiques. Les complexes thérapeutiques préférés ou conjugués comprennent un acide gras oméga 3, tel que l'acide alpha-linolénique, l'acide eicosapentaénoïque ou l'acide docosahexaénoïque et leurs dérivés.
PCT/US1995/009870 1994-08-05 1995-08-04 Complexes biomoleculaires diriges Ceased WO1996004001A1 (fr)

Priority Applications (2)

Application Number Priority Date Filing Date Title
EP95929378A EP0952841A4 (fr) 1994-08-05 1995-08-04 Complexes biomoleculaires diriges
AU32755/95A AU3275595A (en) 1994-08-05 1995-08-04 Site-specific biomolecular complexes

Applications Claiming Priority (4)

Application Number Priority Date Filing Date Title
US08/286,327 1994-08-05
US08/286,327 US5716614A (en) 1994-08-05 1994-08-05 Method for delivering active agents to mammalian brains in a complex with eicosapentaenoic acid or docosahexaenoic acid-conjugated polycationic carrier
US08/487,693 1995-06-07
US08/487,693 US6005004A (en) 1994-08-05 1995-06-07 Lipophilic-polycationic delivery systems

Publications (1)

Publication Number Publication Date
WO1996004001A1 true WO1996004001A1 (fr) 1996-02-15

Family

ID=26963733

Family Applications (1)

Application Number Title Priority Date Filing Date
PCT/US1995/009870 Ceased WO1996004001A1 (fr) 1994-08-05 1995-08-04 Complexes biomoleculaires diriges

Country Status (3)

Country Link
EP (1) EP0952841A4 (fr)
AU (1) AU3275595A (fr)
WO (1) WO1996004001A1 (fr)

Cited By (44)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO1997020579A3 (fr) * 1995-12-05 1997-09-25 Smithkline Beecham Plc Nouveaux composes et leur utilisation
WO1997044336A1 (fr) * 1996-05-22 1997-11-27 Neuromedica, Inc. Conjugues d'acide cis-docosahexaenoique et de paclitaxel
WO1997044063A3 (fr) * 1996-05-22 1998-02-26 Neuromedica Inc Conjugues d'acide cis-docosahexanoïque et d'agents pharmaceutiques
WO1998017325A3 (fr) * 1996-10-25 1998-06-04 Univ Oregon Health Sciences Conjugues lipidiques, polaires et covalents utilises pour le ciblage de medicaments
WO1999004824A1 (fr) * 1997-07-25 1999-02-04 Sdg, Inc. Compositions pharmaceutiques a base de polymeres destinees a une administration ciblee d'agents biologiquement actifs
WO1999008711A1 (fr) * 1997-08-13 1999-02-25 Chiron Corporation Composes de lipides et de polyamides conjugees et compositions pour l'administration d'acides nucleiques
US6063759A (en) * 1990-11-01 2000-05-16 Oregon Health Sciences University Conjugate of biologically active compound and polar lipid conjugated to a microparticle for biological targeting
EP1003771A1 (fr) * 1997-08-12 2000-05-31 Synt:em (S.A.) Peptides lineaires derives de peptides antibiotiques, leur preparation et leur utilisation pour vectoriser des substances actives
US6197764B1 (en) 1997-11-26 2001-03-06 Protarga, Inc. Clozapine compositions and uses thereof
US6225444B1 (en) 1998-02-10 2001-05-01 Protarga, Inc. Neuroprotective peptides and uses thereof
US6258836B1 (en) 1988-02-26 2001-07-10 Protarga, Inc. Dopamine analog amide
WO2001045744A3 (fr) * 1999-12-21 2002-05-02 Ramchand Chaniyilparampu Nanap Nouveaux composes d'acides gras non satures
US6576254B1 (en) * 1998-05-23 2003-06-10 University Of Strathclyde Polyamino acid vesicles
WO2002098455A3 (fr) * 2001-06-07 2004-03-25 Hemebiotech As Production de l'enzyme arylsulfatase a humaine recombinee
US6919076B1 (en) 1998-01-20 2005-07-19 Pericor Science, Inc. Conjugates of agents and transglutaminase substrate linking molecules
US7045543B2 (en) 2001-11-05 2006-05-16 Enzrel Inc. Covalent conjugates of biologically-active compounds with amino acids and amino acid derivatives for targeting to physiologically-protected sites
US7235583B1 (en) 1999-03-09 2007-06-26 Luitpold Pharmaceuticals, Inc., Fatty acid-anticancer conjugates and uses thereof
US7244764B2 (en) 2003-06-23 2007-07-17 Neurochem (International) Limited Methods and compositions for treating amyloid-related diseases
JP2008509229A (ja) * 2004-03-02 2008-03-27 イッサム・リサーチ・ディベロップメント・カンパニー・オブ・ザ・ヘブルー・ユニバーシティ・オブ・エルサレム 疾病の治療における脂質複合体の使用
US7414076B2 (en) 2003-06-23 2008-08-19 Neurochem (International) Limited Methods and compositions for treating amyloid-related diseases
EP2042203A2 (fr) 2007-09-26 2009-04-01 Aesculap AG Prothèse vasculaire renforcée ayant un effet antimicrobien prolongé
WO2010035261A2 (fr) 2008-09-29 2010-04-01 Ben Gurion University Of The Negev Research And Development Authority Beta-peptides amyloides et procédés d'utilisation associés
US7772196B2 (en) 2000-01-10 2010-08-10 Yissum Research Development Company Of The Hebrew University Of Jerusalem Use of lipid conjugates in the treatment of diseases
US7811999B2 (en) 2000-01-10 2010-10-12 Yissum Research Development Company Of The Hebrew University Of Jerusalem, Ltd. Use of lipid conjugates in the treatment of diseases
US7893226B2 (en) 2004-09-29 2011-02-22 Yissum Research Development Company Of The Hebrew University Of Jerusalem, Ltd. Use of lipid conjugates in the treatment of diseases
US8076312B2 (en) 2000-01-10 2011-12-13 Yissum Research Development Company Of The Hebrew University Of Jerusalem Ltd Use of lipid conjugates in the treatment of disease
US8304395B2 (en) 2000-01-10 2012-11-06 Yissum Research Development Company Of The Hebrew University Of Jerusalem Ltd. Lipid conjugates in the treatment of disease
US8372815B2 (en) 2000-01-10 2013-02-12 Yissum Research Development Company Use of lipid conjugates in the treatment of conjunctivitis
US8501701B2 (en) 2000-01-10 2013-08-06 Yissum Research Development Company Of The Hebrew University Of Jerusalem Ltd. Use of lipid conjugates in the treatment of disease
US8663631B2 (en) 2010-09-09 2014-03-04 Synageva Biopharma Corporation Methods for treating lysosomal acid lipase deficiency in patients
US8835654B2 (en) 2004-12-22 2014-09-16 Bhi Limited Partnership Method and compositions for treating amyloid-related diseases
US8859524B2 (en) 2005-11-17 2014-10-14 Yissum Research Development Company Of The Hebrew University Of Jerusalem Lipid conjugates in the treatment of chronic rhinosinusitis
US8865681B2 (en) 2004-03-02 2014-10-21 Yissum Research Development Company of the Hebrew Unitersity of Jerusalem Use of lipid conjugates in the treatment of diseases or disorders of the eye
US8883761B2 (en) 2001-01-10 2014-11-11 Yissum Research Development Company Of The Hebrew University Of Jerusalem Use of lipid conjugates in the treatment of diseases associated with vasculature
US8906882B2 (en) 2005-11-17 2014-12-09 Yissum Research Development Company Of The Hebrew University Of Jerusalem Lipid conjugates in the treatment of allergic rhinitis
US8916539B2 (en) 2000-01-10 2014-12-23 Yissum Research Development Company Of The Hebrew University Of Jerusalem Use of lipid conjugates in the treatment of disease
US8945542B2 (en) 2011-02-15 2015-02-03 Synageva Biopharma Corp. Methods for treating lysosomal acid lipase deficiency
US9040078B2 (en) 2000-01-10 2015-05-26 Yissum Research Development Company Of The Hebrew University Of Jerusalem Use of lipid conjugates in the treatment of diseases of the nervous system
US9499480B2 (en) 2006-10-12 2016-11-22 Bhi Limited Partnership Methods, compounds, compositions and vehicles for delivering 3-amino-1-propanesulfonic acid
WO2018226992A1 (fr) 2017-06-07 2018-12-13 Adrx, Inc. Inhibiteur d'agrégation de tau
WO2019036725A2 (fr) 2017-08-18 2019-02-21 Adrx, Inc. Inhibiteurs peptidiques d'agrégation de tau
US10407671B2 (en) 2010-04-23 2019-09-10 Alexion Pharmaceuticals, Inc. Lysosomal storage disease enzymes
US10864255B2 (en) 2000-02-04 2020-12-15 Children's Hospital Medical Center Lipid hydrolysis therapy for atherosclerosis and related diseases
US11013811B2 (en) 2009-05-11 2021-05-25 Yissum Research Development Company Of The Hebrew University Of Jerusalem Ltd. Lipid-polymer conjugates, their preparation and uses thereof

Citations (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO1992020362A1 (fr) * 1991-05-14 1992-11-26 Shashoua Victor E Peptides favorisant le developpement de cellules nerveuses et leurs utilisations

Family Cites Families (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
ATE138686T1 (de) * 1988-12-23 1996-06-15 Genzyme Corp Cho-zellen befähigt enzymatisch aktive rekombinante-glukocerebrosidase zu produzieren
AU5743994A (en) * 1992-12-10 1994-07-04 Enzon, Inc. Glycolipid enzyme-polymer conjugates

Patent Citations (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO1992020362A1 (fr) * 1991-05-14 1992-11-26 Shashoua Victor E Peptides favorisant le developpement de cellules nerveuses et leurs utilisations

Non-Patent Citations (3)

* Cited by examiner, † Cited by third party
Title
CLINICAL AND EXPERIMENTAL METASTASIS, Volume 11, Number 2, issued 1993, T. NARAI et al., "Intravenously Injected Radiolabelled Fatty Acids Image Brain Tumour Phospholipids in Vivo: Differential Uptakes of Palmitate, Arachidonate and Docosahexaenoate", pages 141-149. *
NEUROSCIENCE LETTERS, Volume 171, issued 1994, M. ALBERGHINA et al., "Differential Transport of Docosahexaenoate and Palmitate Through the Blood-Retina and Blood-Brain Barrier of the Rat", pages 133-136. *
See also references of EP0952841A4 *

Cited By (76)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US6258836B1 (en) 1988-02-26 2001-07-10 Protarga, Inc. Dopamine analog amide
US6407137B2 (en) 1988-02-26 2002-06-18 Protarga, Inc. Dopamine analog amide
US6063759A (en) * 1990-11-01 2000-05-16 Oregon Health Sciences University Conjugate of biologically active compound and polar lipid conjugated to a microparticle for biological targeting
US6339060B1 (en) 1990-11-01 2002-01-15 Oregon Health & Science University Conjugate of biologically active compound and polar lipid conjugated to a microparticle for biological targeting
US5827819A (en) * 1990-11-01 1998-10-27 Oregon Health Sciences University Covalent polar lipid conjugates with neurologically active compounds for targeting
US6858582B2 (en) 1990-11-01 2005-02-22 Oregon Health And Sciences University Composition containing porous microparticle impregnated with biologically-active compound for treatment of infection
US6024977A (en) * 1990-11-01 2000-02-15 Oregon Health Sciences University Covalent polar lipid conjugates with neurologically active compounds for targeting
US6436437B1 (en) 1990-11-01 2002-08-20 Oregon Health And Science University Covalent polar lipid conjugates with neurologically active compounds for targeting
US7423010B2 (en) 1994-05-19 2008-09-09 Oregon Health & Science University Nonporous microparticle-prodrug conjugates for treatment of infection
WO1997020579A3 (fr) * 1995-12-05 1997-09-25 Smithkline Beecham Plc Nouveaux composes et leur utilisation
EP1466628A1 (fr) * 1996-05-22 2004-10-13 Luitpold Pharmaceuticals, Inc. Conjugués DHA et d'un agent pharmaceutique.
US7199151B2 (en) 1996-05-22 2007-04-03 Luitpold Pharmaceuticals, Inc. DHA-pharmaceutical agent conjugates of taxanes
WO1997044063A3 (fr) * 1996-05-22 1998-02-26 Neuromedica Inc Conjugues d'acide cis-docosahexanoïque et d'agents pharmaceutiques
WO1997044336A1 (fr) * 1996-05-22 1997-11-27 Neuromedica, Inc. Conjugues d'acide cis-docosahexaenoique et de paclitaxel
WO1998017325A3 (fr) * 1996-10-25 1998-06-04 Univ Oregon Health Sciences Conjugues lipidiques, polaires et covalents utilises pour le ciblage de medicaments
WO1999004824A1 (fr) * 1997-07-25 1999-02-04 Sdg, Inc. Compositions pharmaceutiques a base de polymeres destinees a une administration ciblee d'agents biologiquement actifs
EP1003771A1 (fr) * 1997-08-12 2000-05-31 Synt:em (S.A.) Peptides lineaires derives de peptides antibiotiques, leur preparation et leur utilisation pour vectoriser des substances actives
WO1999008711A1 (fr) * 1997-08-13 1999-02-25 Chiron Corporation Composes de lipides et de polyamides conjugees et compositions pour l'administration d'acides nucleiques
US6569450B1 (en) 1997-08-13 2003-05-27 Chiron Corporation Lipid-conjugated polyamide compounds and related compositions and methods thereof
US6572881B1 (en) 1997-08-13 2003-06-03 Chiron Corporation Lipid-conjugated polyamide compounds and related compositions and methods thereof
US7214384B2 (en) 1997-08-13 2007-05-08 Novartis Vaccines And Diagnostics, Inc. Lipid-conjugated polyamide compounds
US6197332B1 (en) 1997-08-13 2001-03-06 Chiron Corporation Lipid-conjugated polyamide compounds and related compositions and methods thereof
US6197764B1 (en) 1997-11-26 2001-03-06 Protarga, Inc. Clozapine compositions and uses thereof
US6919076B1 (en) 1998-01-20 2005-07-19 Pericor Science, Inc. Conjugates of agents and transglutaminase substrate linking molecules
US6225444B1 (en) 1998-02-10 2001-05-01 Protarga, Inc. Neuroprotective peptides and uses thereof
US6627601B2 (en) 1998-02-10 2003-09-30 Protarga, Inc. Neuroprotective peptides and uses thereof
US6576254B1 (en) * 1998-05-23 2003-06-10 University Of Strathclyde Polyamino acid vesicles
US7235583B1 (en) 1999-03-09 2007-06-26 Luitpold Pharmaceuticals, Inc., Fatty acid-anticancer conjugates and uses thereof
WO2001045744A3 (fr) * 1999-12-21 2002-05-02 Ramchand Chaniyilparampu Nanap Nouveaux composes d'acides gras non satures
US7772196B2 (en) 2000-01-10 2010-08-10 Yissum Research Development Company Of The Hebrew University Of Jerusalem Use of lipid conjugates in the treatment of diseases
US8501701B2 (en) 2000-01-10 2013-08-06 Yissum Research Development Company Of The Hebrew University Of Jerusalem Ltd. Use of lipid conjugates in the treatment of disease
US8383787B2 (en) 2000-01-10 2013-02-26 Yissum Research Development Company Use of lipid conjugates in the treatment of diseases
US8372815B2 (en) 2000-01-10 2013-02-12 Yissum Research Development Company Use of lipid conjugates in the treatment of conjunctivitis
US8304395B2 (en) 2000-01-10 2012-11-06 Yissum Research Development Company Of The Hebrew University Of Jerusalem Ltd. Lipid conjugates in the treatment of disease
US9040078B2 (en) 2000-01-10 2015-05-26 Yissum Research Development Company Of The Hebrew University Of Jerusalem Use of lipid conjugates in the treatment of diseases of the nervous system
US9012396B2 (en) 2000-01-10 2015-04-21 Yissum Research Development Company Of The Hebrew University Of Jerusalem Use of lipid conjugates in the treatment of conjunctivitis
US8916539B2 (en) 2000-01-10 2014-12-23 Yissum Research Development Company Of The Hebrew University Of Jerusalem Use of lipid conjugates in the treatment of disease
US8865878B2 (en) 2000-01-10 2014-10-21 Yissum Research Development Company Of The Hebrew University Of Jerusalem Use of lipid conjugates in the treatment of diseases
US8076312B2 (en) 2000-01-10 2011-12-13 Yissum Research Development Company Of The Hebrew University Of Jerusalem Ltd Use of lipid conjugates in the treatment of disease
US8901103B2 (en) 2000-01-10 2014-12-02 Yissum Research Development Company Of The Hebrew University Of Jerusalem Use of lipid conjugates in the treatment of diseases
US7811999B2 (en) 2000-01-10 2010-10-12 Yissum Research Development Company Of The Hebrew University Of Jerusalem, Ltd. Use of lipid conjugates in the treatment of diseases
US10864255B2 (en) 2000-02-04 2020-12-15 Children's Hospital Medical Center Lipid hydrolysis therapy for atherosclerosis and related diseases
US8883761B2 (en) 2001-01-10 2014-11-11 Yissum Research Development Company Of The Hebrew University Of Jerusalem Use of lipid conjugates in the treatment of diseases associated with vasculature
WO2002098455A3 (fr) * 2001-06-07 2004-03-25 Hemebiotech As Production de l'enzyme arylsulfatase a humaine recombinee
US7045543B2 (en) 2001-11-05 2006-05-16 Enzrel Inc. Covalent conjugates of biologically-active compounds with amino acids and amino acid derivatives for targeting to physiologically-protected sites
US7244764B2 (en) 2003-06-23 2007-07-17 Neurochem (International) Limited Methods and compositions for treating amyloid-related diseases
US7598269B2 (en) 2003-06-23 2009-10-06 Bellus Health (International) Limited Methods and compositions for treating amyloid-related diseases
US7414076B2 (en) 2003-06-23 2008-08-19 Neurochem (International) Limited Methods and compositions for treating amyloid-related diseases
EP1758595A4 (fr) * 2004-03-02 2009-12-16 Yissum Res Dev Co Utilisation de conjugues lipidiques dans le traitement de maladies
US8865681B2 (en) 2004-03-02 2014-10-21 Yissum Research Development Company of the Hebrew Unitersity of Jerusalem Use of lipid conjugates in the treatment of diseases or disorders of the eye
JP2012092341A (ja) * 2004-03-02 2012-05-17 Yissum Research Development Co Of The Hebrew Univ Of Jerusalem Ltd 疾病の治療における脂質複合体の使用
JP2008509229A (ja) * 2004-03-02 2008-03-27 イッサム・リサーチ・ディベロップメント・カンパニー・オブ・ザ・ヘブルー・ユニバーシティ・オブ・エルサレム 疾病の治療における脂質複合体の使用
US7893226B2 (en) 2004-09-29 2011-02-22 Yissum Research Development Company Of The Hebrew University Of Jerusalem, Ltd. Use of lipid conjugates in the treatment of diseases
US8835654B2 (en) 2004-12-22 2014-09-16 Bhi Limited Partnership Method and compositions for treating amyloid-related diseases
US8906882B2 (en) 2005-11-17 2014-12-09 Yissum Research Development Company Of The Hebrew University Of Jerusalem Lipid conjugates in the treatment of allergic rhinitis
US8859524B2 (en) 2005-11-17 2014-10-14 Yissum Research Development Company Of The Hebrew University Of Jerusalem Lipid conjugates in the treatment of chronic rhinosinusitis
US11020360B2 (en) 2006-10-12 2021-06-01 Bellus Health Inc. Methods, compounds, compositions and vehicles for delivering 3-amino-1-propanesulfonic acid
US10238611B2 (en) 2006-10-12 2019-03-26 Bellus Health Inc. Methods, compounds, compositions and vehicles for delivering 3-amino-1-propanesulfonic acid
US9499480B2 (en) 2006-10-12 2016-11-22 Bhi Limited Partnership Methods, compounds, compositions and vehicles for delivering 3-amino-1-propanesulfonic acid
US10857109B2 (en) 2006-10-12 2020-12-08 Bellus Health, Inc. Methods, compounds, compositions and vehicles for delivering 3-amino-1-propanesulfonic acid
DE202008017254U1 (de) 2007-09-26 2009-04-30 Aesculap Ag Verstärkte Gefäßprothese mit antimikrobieller Langzeitwirkung
US8940039B2 (en) 2007-09-26 2015-01-27 Aesculap Ag Reinforced vascular prosthesis with long-term antimicrobial action
DE102007047246A1 (de) 2007-09-26 2009-04-02 Aesculap Ag Verstärkte Gefäßprothese mit antimikrobieller Langzeitwirkung
EP2042203A2 (fr) 2007-09-26 2009-04-01 Aesculap AG Prothèse vasculaire renforcée ayant un effet antimicrobien prolongé
WO2010035261A2 (fr) 2008-09-29 2010-04-01 Ben Gurion University Of The Negev Research And Development Authority Beta-peptides amyloides et procédés d'utilisation associés
US11013811B2 (en) 2009-05-11 2021-05-25 Yissum Research Development Company Of The Hebrew University Of Jerusalem Ltd. Lipid-polymer conjugates, their preparation and uses thereof
US10407671B2 (en) 2010-04-23 2019-09-10 Alexion Pharmaceuticals, Inc. Lysosomal storage disease enzymes
US10858638B2 (en) 2010-04-23 2020-12-08 Alexion Pharmaceuticals, Inc. Lysosomal storage disease enzymes
US11560554B2 (en) 2010-04-23 2023-01-24 Alexion Pharmaceuticals, Inc. Lysosomal storage disease enzymes
US10166274B2 (en) 2010-09-09 2019-01-01 Alexion Pharmaceuticals, Inc. Methods for treating lysosomal acid lipase deficiency in patients
US8663631B2 (en) 2010-09-09 2014-03-04 Synageva Biopharma Corporation Methods for treating lysosomal acid lipase deficiency in patients
US11400141B2 (en) 2010-09-09 2022-08-02 Alexion Pharmaceuticals, Inc. Methods for treating lysosomal acid lipase deficiency in patients
US12076376B2 (en) 2010-09-09 2024-09-03 Alexion Pharmaceuticals, Inc. Methods for treating lysosomal acid lipase deficiency in patients
US8945542B2 (en) 2011-02-15 2015-02-03 Synageva Biopharma Corp. Methods for treating lysosomal acid lipase deficiency
WO2018226992A1 (fr) 2017-06-07 2018-12-13 Adrx, Inc. Inhibiteur d'agrégation de tau
WO2019036725A2 (fr) 2017-08-18 2019-02-21 Adrx, Inc. Inhibiteurs peptidiques d'agrégation de tau

Also Published As

Publication number Publication date
EP0952841A1 (fr) 1999-11-03
EP0952841A4 (fr) 2000-11-02
AU3275595A (en) 1996-03-04

Similar Documents

Publication Publication Date Title
US6005004A (en) Lipophilic-polycationic delivery systems
WO1996004001A1 (fr) Complexes biomoleculaires diriges
Vyas et al. Ligand-receptor-mediated drug delivery: an emerging paradigm in cellular drug targeting
Lv et al. Toxicity of cationic lipids and cationic polymers in gene delivery
Lalatsa et al. Strategies to deliver peptide drugs to the brain
KR101186210B1 (ko) 혈뇌장벽을 통과하는 물질 수송용 인공 저밀도 지단백질 운반체
US6221397B1 (en) Surface cross-linked particles suitable for controlled delivery
US5583020A (en) Permeability enhancers for negatively charged polynucleotides
US6312727B1 (en) Delivery of nucleic acid materials
US6030941A (en) Polymer composition for delivering substances in living organisms
US20110014235A1 (en) Therapeutic and vaccine polyelectrolyte nanoparticle compositions
CA2150956A1 (fr) Conjugues utiles contre les glycolipides et certains polymeres
CA2506842A1 (fr) Amplification du ciblage medie par la biotine
CA2372834A1 (fr) Amplification du ciblage a mediation folate de cellules tumorales a l'aide de nanoparticules
Zhang et al. A brief review of polysialic acid-based drug delivery systems
Lee et al. Folate-targeted liposomes for drug delivery
CA2492862A1 (fr) Administration par voie orale d'un agent therapeutique couple a un agent de transport
Fujita et al. Control of in vivo fate of albumin derivatives utilizing combined chemical modification
Blanchfield et al. Lipid, sugar and liposaccharide based delivery systems 2
Krinick et al. Soluble polymers as targetable drug carriers
Ali et al. Peptide delivery systems
US20050220754A1 (en) Vitamin directed targeting therapy
Gagliardi Recent advances in preclinical studies and potential applications of dendrimers as drug carriers in the central nervous system
Poznansky Enzyme—Albumin polymers: New approaches to the use of enzymes in medicine
Ahmad et al. Strategies for designing antibody-toxin conjugates

Legal Events

Date Code Title Description
AK Designated states

Kind code of ref document: A1

Designated state(s): AU CA HU JP NO

AL Designated countries for regional patents

Kind code of ref document: A1

Designated state(s): AT BE CH DE DK ES FR GB GR IE IT LU MC NL PT SE

DFPE Request for preliminary examination filed prior to expiration of 19th month from priority date (pct application filed before 20040101)
121 Ep: the epo has been informed by wipo that ep was designated in this application
WWE Wipo information: entry into national phase

Ref document number: 1995929378

Country of ref document: EP

NENP Non-entry into the national phase

Ref country code: CA

WWP Wipo information: published in national office

Ref document number: 1995929378

Country of ref document: EP

WWW Wipo information: withdrawn in national office

Ref document number: 1995929378

Country of ref document: EP