EP1370138A1 - Therapie genique pour tissus neurologiques - Google Patents

Therapie genique pour tissus neurologiques

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Publication number
EP1370138A1
EP1370138A1 EP02713697A EP02713697A EP1370138A1 EP 1370138 A1 EP1370138 A1 EP 1370138A1 EP 02713697 A EP02713697 A EP 02713697A EP 02713697 A EP02713697 A EP 02713697A EP 1370138 A1 EP1370138 A1 EP 1370138A1
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EP
European Patent Office
Prior art keywords
cerebrospinal
flow
growth factor
gene therapy
fluid
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Withdrawn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
EP02713697A
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German (de)
English (en)
Other versions
EP1370138A4 (fr
Inventor
David P. Hesson
Glenn D. Frazer
Bruce Shook
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.)
Neuron Therapeutics Inc
Original Assignee
Neuron Therapeutics Inc
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Publication date
Application filed by Neuron Therapeutics Inc filed Critical Neuron Therapeutics Inc
Publication of EP1370138A1 publication Critical patent/EP1370138A1/fr
Publication of EP1370138A4 publication Critical patent/EP1370138A4/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Withdrawn legal-status Critical Current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
    • A61K48/00—Medicinal preparations containing genetic material which is inserted into cells of the living body to treat genetic diseases; Gene therapy
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P21/00—Drugs for disorders of the muscular or neuromuscular system
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P25/00—Drugs for disorders of the nervous system
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P3/00—Drugs for disorders of the metabolism

Definitions

  • This invention relates to gene therapy formulations or compositions and methods useful for conducting gene therapy for neurological tissue, of the central nervous system (CNS).
  • the compositions can contain an artificial cerebrospinal fluid (ACSF) carrier component and can further contain nutrients.
  • ACSF cerebrospinal fluid
  • the cerebrospinal fluid (CSF) pathway system which intimately bathes and permeates brain and spinal cord tissues, constitutes a circulatory system within the body. Although it has some similarities to systemic vascular and lymphatic circulation, its anatomical arrangement differs considerably. Indeed, this system has been named the “third circulation” system. Due to the extensive area of CSF-tissue contact over the cerebral and spinal cord surfaces, in the paravascular Virchow-Robins spaces, and cerebral ventricles, the cerebrospinal fluid system constitutes a vast, complex and intimate avenue for access to central nervous tissue.
  • nucleic acid construct that expresses an antisense RNA to interfere in the expression of a certain mRNA or one or more constructs that express two complementary strands designed to interfere in the expression of a certain mRNA.
  • nucleic acid-based vaccines seek to induce a percentage of cells to produce immune-reaction inducing polypeptides, to induce an antibody-based or cellular-based immune response. Also, one can seek to inhibit the function of genes or gene products with antisense molecules.
  • the present invention provides efficient avenues for delivering gene therapy agents to neuronal tissue.
  • the invention provides a method of delivering to neurological tissue of the CNS a transforming composition or a transformed cell, the method comprising: a. injecting a physiologically acceptable cerebrospinal perfusion fluid (CSPF) into a first catheter into the cerebrospinal pathway, which CSPF has a gene therapy effective amount of gene therapy agent or transformed cells; b. withdrawing fluid at a second catheter into the cerebrospinal pathway to create a flow and flow pathway between the first and second catheters; and c. maintaining the flow for a period of time adapted to perfuse at least 1 CSF volume.
  • CSPF cerebrospinal perfusion fluid
  • the invention provides a method of delivering to neurological tissue a transforming composition or a transformed cell, the method comprising: a.
  • cerebrospinal flushing fluid injecting a physiologically acceptable cerebrospinal flushing fluid into a first catheter into the cerebrospinal pathway, which cerebrospinal flushing fluid has an effective amount a gene therapy agent effective to transform neural cells to express calbindin, bal-2 leptin superoxide dismutase, a glial-derived neurotrophic growth factor which is NGF, BDNF or NT3, epidermal growth factor, a fibroblast growth factor which is FGF 1 -9, insulin-like growth factor- 1, a platelet-derived growth factor which is PDGF- A, B or C, vascular endothelial growth factor, or ciliary neurotrophic factor; or transformed cells; b.
  • a gene therapy agent effective to transform neural cells to express calbindin, bal-2 leptin superoxide dismutase, a glial-derived neurotrophic growth factor which is NGF, BDNF or NT3, epidermal growth factor, a fibroblast growth factor which is F
  • the invention provides a method of delivering to neurological tissue a transforming composition or a transformed cell, the method comprising: a. injecting a cerebrospinal perfusion fluid into a first catheter into the cerebrospinal pathway, which cerebrospinal perfusion fluid has a gene therapy effective amount of transforming nucleic acid, wherein the cerebrospinal flushing fluid further comprises an emulsion-forming effective amount of a lipid composition comprised of lipids found in biological membranes; b. withdrawing fluid at a second catheter into the cerebrospinal pathway to create a flow and flow pathway between the first and second catheters; and c. maintaining the flow for a period of time adapted to flush at least 1 CSF volume.
  • Figure 1 illustrates a perfusion pathway.
  • Vectors typically transduce only a percentage of the cells to which they are applied.
  • the transducing gene is often maintained on an episome and is therefore often not a stably incorporated and maintained genetic element.
  • incorporation into the chromosomal DNA is often dependent on cell division, thereby limiting the scope of target tissues to replicating tissues.
  • Viral vectors can encode proteins that induce immunity, thereby carrying the seeds for the destruction of the transduced cells. Certain viral vectors overcome some of these problems but otherwise create at least an implication of danger.
  • non-replicating forms of the human immunodeficiency virus are being engineered for use as gene therapy vectors that allow for the incorporation of the genetic material into genomic DNA.
  • nucleic acid-based vaccines for example, an immune response is desirable, as can be a process by which expression of the transforming gene attenuates so that production of the immuno-stimulants attenuates over time.
  • Viral vectors have also been subject to engineering to change their target cell preference, for instance by binding or incorporating antibodies.
  • Valsesia- Wittmann et al. modified the cell-surface binding characteristics of avian leucosis virus (J. Virol. 68: 4609-4619, 1994).
  • Erythropoietin which of course binds its cognate receptor, has been incorporated into Moloney murine leukemia virus (Mo-MLV) (Kasahara et al., Science 266: 1373-1376, 1994).
  • a tumor- targeting single-chain antibody has been incorporated into spleen necrosis virus (Chu and Dornburg, J. Virol. 69: 2659-2663, 1995).
  • HIV envelope protein has been incorporated into murine leukemia viral vectors (Mammamo et al., J. Virol. 71 : 3341 -3345, 1997).
  • Such targeting methods with respect to adenoviral vectors are reviewed by Reynolds and Curiel ("Strategies to Adapt Adenoviral Vectors for Gene Therapy Applications: Targeting and Integration," in Development of Human Gene Therapy, Cold Spring Harbor Laboratory Press, 1998, pp. 1 1 1-130).
  • Another approach with adenoviral vectors is to make hybrid vectors using the viral coat protein from an adenovirus strain with an appropriate target specificity.
  • a vector from Cobra Therapeutics is based on the Ad5 strain, but uses the coat protein from Ad35 to obtain a vector effective with dendritic cells.
  • a wide variety of viral vectors have been selected or engineered for gene therapy.
  • nucleic acid can also be delivered successfully without the use of viral vectors.
  • compositions that have been used for non- virally mediated transfection will prove more appropriate for use to transform cells, which cells can be introduced into neurological tissues by the methods of the invention.
  • Such methods include, for example, an early-developed method for increasing transfection efficiency was to use calcium phosphate-precipitated nucleic acid.
  • the transfection potential of nucleic acid is increased by compacting it with polycationic polymers such as DEAE dextran (Veheri et al., Virology 27: 434-436, 1965), polylysine (Wu et al., J. Biol. Chem. 266: 14338-14342 1991), cationic peptides (Wadhwa et al., Bioconjugate Chem.
  • enhancers of nucleic acid uptake include erodable microspheres (Mathiowitz et al., Nature 386: 410-412, 1997) and polyvinyl pyrrolidone (Mumper et al., Pharm. Res. 13: 701-709, 1996).
  • enhancers include cationic liposomes into which the nucleic acid is incorporated (Feigner et al., Proc. Natl. Acad. Sci. U.S.A. 84:7413-7, 1987; Feigner and Ringold, Nature 337:387-8, 1989).
  • liposomes are believed to insert the nucleic acid into a target cell by a membrane fusion mechanism.
  • Illustrative of the many cationic lipid formulations now available is DOTMA (N[l-(2,3-dioleyloxy)propyl]-NNN-trimethylammonium).
  • cationic lipid formulations include Lipofectin I M , a 1 :1 (w/w) liposome formulation of the cationic lipid ⁇ -[l-(2,3-dioleyloxy)propyl]- ⁇ , ⁇ , ⁇ - trimethylammonium chloride (DOTMA) and dioleoyl phosphatidylethanolamine (DOPE), LipofectAMINETM, a 3: 1 (w/w) liposome formulation of the polycationic lipid 2,3-dioleyloxy-N-[2(spermine-carboxamido)ethyl]-N,N-dimethyl-l- propanaminiumtrifluoroacetate (DOSPA) and the neutral lipid dioleoyl phosphatidylethanolamine (DOPE) in membrane-filtered water, and LipofectACE M , a 1 :2.5 (w/w) liposome formulation of the cationic lipid dimethyl dioctadecylammonium bro
  • Non-viral techniques that are more preferred for direct use in neurological tissue include erodible microspheres and polyvinyl pyrrolidone mediated techniques, as well as techniques that utilize liposomal compositions enriched in natural cationic lipids. Also preferred are gene transfers effected without such adjuvants.
  • Targeting techniques can also be employed which bind or affix targeting molecules to the nucleic acid or nucleic acid complex to be used for transfection.
  • Cotton and Wagner "Receptor-mediated Gene Delivery Strategies," in Development of Human Gene Therapy, Cold Spring Harbor Laboratory Press, 1998, pp. 261-277.
  • Neuronal tissue of the CNS is to some degree immunologically privileged. The population of immune cells present is typically lower than in other tissues.
  • vaccinations by the methods of the invention are conducted as an adjunct to a prophylactic or therapeutic vaccination made by another route of administration.
  • Gene therapy approaches seeking to induce cellular immunity can also be conducted by the methods of the invention.
  • the invention further encompasses using suicide bacteria (attenuated, intracellular parasitic bacteria) to deliver nucleic acid to the cytoplasm of bacteria- ingesting cells such as macrophages. See, e.g., US Patent 6,143,551.
  • the invention also encompasses delivering cells that were transformed in vitro.
  • Such cells can include, for example, pluripotent cells such as fetal brain cells.
  • pluripotent cells such as fetal brain cells.
  • bone marrow cells, other primary stem cells or stem cell lines, such as F9 cells which have recently been shown to include cells capable of differentiating to neurological cells (Mezey et al., Science 290: 1779-1782, Dec. 1, 2000; Brazelton et al., .Science 290:1775-1779, Dec. 1, 2000).
  • the transfection-facilitating adjuvant is the emulsified lipid component of cerebrospinal perfusion fluid (described below).
  • the nucleic acid sought to be introduced into cells will often include, in addition to the portion conveying the primary genetic characteristic of interest, a portion encoding a substance that is itself, or gives rise to, a molecule that is readily detectable.
  • This "reporter" molecule serves as a surrogate for determining or estimating success in introducing the primary genetic characteristic.
  • a portion of the nucleic acid can encode a substance required for the cells to survive in the face of an appropriate challenge.
  • the nucleic acid can be single or double-stranded, though non-virally mediated techniques that seek to express a portion of the nucleic acid will typically use double-stranded nucleic acid.
  • antisense molecules such as antisense oligonucleotides
  • hybridization arrest an antisense molecule inhibitor binds to the target nucleic acid and thus prevents its utilization, for example by sterically hindering the binding of essential proteins, such as ribosomes, to the nucleic acid.
  • Methyl phosphonate antisense molecules and ⁇ -anomer antisense molecules are two extensively studied antisense agents which are thought to disrupt nucleic acid function by hybridization arrest (Miller and Ts'O, Anti-Cancer Drug Design, 2:117-128, 1987).
  • a 2'-deoxyribofuranosyl antisense molecule or antisense molecule analog hybridizes with the targeted RNA and this duplex activates the RNase H enzyme to cleave the RNA strand.
  • Phosphorothioate antisense molecules are the most prominent example of an antisense agent that is believed to primarily operate by this second mechanism.
  • Antisense molecules can also be adapted to bind duplex nucleic acids to form triplex complexes in a sequence specific manner via Hoogsteen base pairing (Beal et al., Science 251 : 1360-1363, 1991 ; Young et al., Proc. Nail. Acad. Sci. 88: 10023-10026, 1991). Both antisense and triple helix therapeutic strategies are typically directed towards nucleic acid sequences that are involved in or responsible for establishing or maintaining disease conditions.
  • Target nucleic acid sequences can be, for example, found in the genomes of pathogenic organisms including bacteria, yeasts, fungi, protozoa, parasites, viruses, or may be endogenous in nature.
  • Modifications have been made to the ribose phosphate backbone of antisense molecules to increase their resistance to nucleases. These modifications include use of, for example, methyl phosphonate, phosphorothioate and phosphorodithioate linkages, as well as alkylphosphonothioate or arylphosphonothioate linkages (see, U.S. Patent 5,929,226 and P. Dan Cook, "Second Generation Antisense Oligonucleotides: 2'- Modifications", Annual Reports in Medicinal Chemistry, 33 :313-325, AcademicPress 1998).
  • Still further substitutions for phosphodiester linkages between a 2' and 5' position of adjacent nucleosides can be -N(R)-CH 2 CH 2 - -CH 2 -N(R)CH 2 , -CH 2 CH 2 N(R)- - OCH 2 CH 2 - -CH 2 CH 2 O- -OCH 2 S-, -SCH 2 CH 2 - -S(0)CH 2 CH 2 - -S(O) 2 CH 2 CH 2 -, - CH 2 S(O) 2 CH 2 - -CH 2 SCH 2 - -NHC(O)0- -OC(O)NH- -OC(O)N(CH 2 )- or -O- CH 2 -O-, where R is lower alkyl.
  • Antisense molecules and antisense molecule analogs having 2'-O-methoxyethyl-substitutions have also been shown to be antisense inhibitors of gene expression with useful features for in vivo use (Martin, Helv. Chim. Acta 78: 486-504, 1995; Altmann et al., Chimia 50: 168-176, 1996; Altmann et al., Biochem. Soc. Trans. 24: 630-637, 1996; and Altmann et al., Nucleosides 16: 917-926, 1997). Relative to DNA-based oligonucleotides, these antisense molecules display improved RNA affinity and higher nuclease resistance.
  • Chimeric antisense molecules with 2'-0-methoxyethyl-ribonucleoside wings and a central DNA- phosphorothioate window also have been shown to effectively reduce the growth of tumors in animal models at low doses.
  • MOE substituted antisense molecules have shown outstanding promise as antisense agents in several disease states. Diseases That Can Be Treated A wide variety of disease and conditions of the brain and spinal cord can be treated with one or a combination of gene therapies.
  • stem cells or proteins such as, calbindin, bcl-2, glial-derived neurotrophic growth factor (NGF, BDNF, NT3), epidermal growth factor (EGF), fibroblast growth factor (FGF 1-9), superoxide dismutase (SOD), insulin (IGF-1), platelet-derived growth factor (PDGF-A, B &C), vascular endothelial growth factor (VEGF), and ciliary neurotrophic factor (CNTF).
  • stem cells or proteins such as, calbindin, bcl-2, glial-derived neurotrophic growth factor (NGF, BDNF, NT3), epidermal growth factor (EGF), fibroblast growth factor (FGF 1-9), superoxide dismutase (SOD), insulin (IGF-1), platelet-derived growth factor (PDGF-A, B &C), vascular endothelial growth factor (VEGF), and ciliary neurotrophic factor (CNTF).
  • NGF glial-derived neurotrophic
  • the cerebrospinal perfusion fluid is an oxygen-carrying [ 0 nutrient emulsion according to the following Table. OXYGEN CARRYING FORMULATIONS:
  • the pH of the emulsion, or vehicle (constituting the above or the like without oxygen-carrying compound), is in the physiological range, such as about pH 7.3.
  • the amino acids include tryptophan.
  • the cerebrospinal perfusion fluid is preferably formulated such that it is physiologic and can directly contact tissues of the neuraxis for an extended period of time, from hours to days, without causing side effects. For best performance, it is believed that the artificial cerebrospinal fluid should be appropriately buffered and have appropriate amounts of amino acids, electrolytes and other compounds helpful to healthy metabolism. Thus, in preferred methods, these components do not need to be supplied through equilibration with other body fluids.
  • cerebrospinal perfusion fluid is formulated with nutrients, it can be termed "artificial cerebrospinal fluid" or "ACSF.”
  • ASF artificial cerebrospinal fluid
  • the cerebrospinal perfusion fluid is simplified further, such as according to the following:
  • the poly-fluorinated, oxygen-carrying compound can be omitted.
  • the amino acid nutrient components can be omitted. Ions are maintained to the degree required to avoid damage to cerebrospinal tissue. Appropriate amounts of oncotic agents are preferred.
  • the cerebrospinal perfusion fluid preferably contains one or both of the lipid and albumin components in the amounts recited above.
  • the lipid can be derived from a lipid source containing lipids of a type used to form biological membranes.
  • the lipids are phospholipids, such as the phospholipid, predominately lecithin, isolated from egg yolk. Where lipids are present, the cerebrospinal perfusion fluid is typically emulsified.
  • tissue and cells will not fare well if exposed to large volumes of non- physiologic ionic solutions. Accordingly, appropriate electrolyte compositions at the tissue level are important when it is considered that the circulatory method of the present invention could dilute of electrolytes from the region, to the detriment of cell membrane function. Desirably, sodium, potassium, calcium, magnesium, and chloride ions are carefully balanced in the antimicrobial formulations of the present invention to create, to the degree possible, normal extra-cellular compositions.
  • the formulations of the invention preferably exclude four amino acids, glutathione, cysteine, ornithine and glutamine, from the group of amino acids included in the formulation, and preferably include sodium bicarbonate in an amount sufficient to increase the buffering capacity of the nutrient solution, in order to more closely resemble cerebrospinal fluid of the subject.
  • Kits for conveniently and safely generating fluorocarbon nutrient emulsion or a corresponding vehicle lacking poly-fluorinated, oxygen-carrying compound are described for example in US Patent Application No. 09/619,414, filed July 19, 2000 (the specific formulations and kits described therein are incorporated by reference as outlined below).
  • the transforming composition is circulated through this cerebrospinal fluid route by injecting it into brain vesicles and withdrawing it from the cisterna magna or the spinal subarachnoid space to nourish and to treat central nervous tissues.
  • the fluid can be injected into the subarachnoid space and withdrawn from another subarachnoid position.
  • the cerebrospinal perfusion fluid comprising a gene therapy agent can be introduced into the subarachnoid spaces through a catheter that transverses the skull or spinal column and the meninges.
  • the delivery point can be the lateral ventricles, subarachnoid space around the brain, cisterna magna or anywhere along the spine.
  • the cerebrospinal perfusion fluid can be withdrawn from the subarachnoid space from any of these locations using a similar catheter.
  • the cerebrospinal perfusion fluid can be returned to the delivery system, reconditioned as necessary to add components that have been consumed or remove undesirable components that have accumulated, and then returned to the subarachnoid space in recirculating fashion. This process can be continued for days if necessary, thereby directly exposing the neuraxis to a gene therapy agent or transformed cells over an extended period of time.
  • This method has several advantages over other routes of administration, such as direct exposure of the nervous system tissue to the gene therapy agent by a simple bolus injection of the agent or cells into the subarachnoid space.
  • This invention provides a method of circulating the gene therapy agent throughout the neuraxis, this exposing nervous system tissue to the agent or cells much more uniformly than would otherwise be possible. It also provides a method of maintaining the gene therapy agent within a narrow concentration range, avoiding the necessity of high concentrations over time. According to this method, the nervous system tissue can be exposed to the agent or cells for extended period time, such as days, if necessary. Further, this method minimizes the amount of agent or cells necessary to achieve a therapeutic effect.
  • a flow pathway from the entry catheter (e.g., a ventricular catheter into a lateral ventricle of the brain) to an exit point at a different location in the cerebral spinal pathway (e.g., into the intrathecal space of the lumbar
  • CSF transforming nucleic acid, oxygen-carrying compound, other emulsified components, or the like.
  • a ventricular catheter 1 is inserted into a lateral ventrical 2. Via aqueduct 3, cisterna magna 4 and subarachnoid spaces 5, a flow pathway can be established to a lumbar outflow catheter 6.
  • the vehicle can be used to establish the existence of a flow pathway (such as that illustrated) from the inflow catheter to the outflow catheter.
  • the vehicle is infused under gravity feed, with the pressure head designed to avoid excessive intracranial pressure.
  • the vehicle can be substituted with the cerebrospinal perfusion fluid.
  • Cerebrospinal perfusion fluid is preferably perfused through the cerebrospinal pathway for a period of time or perfusion volume adapted to effectively presents the gene therapy agent (e.g. nucleic acid or transformed cell).
  • the volume perfused is, in one embodiment, preferably about 15 CSF volumes, where a "CSF volume" is the average volume of CSF fluid found in animals of comparable age to the subject. Preferably, at least about 1 , 2, 4, 8 or 30 CSF volumes are used.
  • a flow rate in the range of 300-3,600 mL/hr is expected, resulting in the exchange of about 2- 22 CSF volumes/hr.
  • the perfusion is preferably with 300 to 3,600 mL/hr.
  • the perfusion can be conducted, for example, for 6, 12, 24 or 48 or more hours. Preferably the perfusion is conducted for between 6 hours and 48 hours or more preferably between 12 hours and 24 hours. More preferably, the perfusion is conducted for at least about 24 hours; and preferably the perfusion is conducted for no more than about 120 hours (and in one embodiment, no more than about 72 hours).
  • Preferred treatment subjects among animals are mammals, preferably humans.
  • Oxy2en-Carryins Compounds Generally, the preferred compounds for use as non-aqueous oxygen transfer components are fluorocarbons, such as perfluorocarbons, perfluorinated alkyl polyethers, fluoroethers, fluoramines, etc. While compounds within these groups range in gram molecular weight from 250 to 7000 g/mole, their selection for use as non-aqueous transport components are based upon the combination of features of the proper vapor pressure, molecular weight, viscosity, ability to form emulsions, emulsion stability and tissue distribution. Not only do fluorocarbons possess appropriate properties but they are for the most part non-toxic.
  • One chief advantage of the CSF circulation route is that most or all of the formulation can be removed by flushing the subarachnoid space with vehicle at the time of treatment termination. In this way long term cellular retention of oxygenating liquids can be avoided.
  • Poly-fluorinated, oxygen-carrying compounds are known in the art. The basic requirement is effectiveness in carrying physiologically useful amounts of oxygen. Factors involved in selecting preferred such compounds include oxygen capacity, tissue retention (preferably minimized), emulsion stability, toxicity, and the like. Such compounds are described in numerous publications (for example, in: Riess et al., "Design Synthesis and Evaluation of Fluorocarbons and Surfactants for In vivo Applications New Perfluoroalkylated Polyhydroxylated Surfactants", Biomat.
  • the double bond is trans.
  • One preferred poly-fluorinated, oxygen-carrying compound is tr ⁇ r ⁇ -Bis-perfluorobutyl ethylene (m and n each equal 4), which is also known as F44E.
  • F44E formulations have a 25% greater oxygen carrying capacity than that of a prior nutrient solution made with perfluorodecalin. Bell et al., Neurology 37: 133, 1987. Formulations comprising F44E are less viscous and relatively easier to perfuse. Also preferred are those of the formula
  • cell-free hemoglobin and liposome encapsulated hemoglobin may also be used as artificial oxygen carriers.
  • Hemoglobin is a 4 subunit protein that is the naturally occurring oxygen carrier in red blood cells.
  • Cell- free hemoglobin rapidly dissociates in the bloodstream, so artificial hemoglobins are chemically modified to prevent breakdown.
  • Artificial hemoglobins can be the product of surface modification, crosslinking, or polymerization. The production and use of cell- free hemoglobin is detailed in a number of publications (for example, U.S. Pat. Nos.
  • Hemoglobin can also be prevented from degradation by being encapsulated within a protective barrier, as in the case with liposome encapsulated hemoglobin, the production and use of which is presented in a number of publications (for example, U.S. Pat. Nos. 5,049,391; 4,133,874; 4,776,991 ; 4,425,334, and 4,532,130).
  • Example 1 Treatment of Amyotrophic Lateral Sclerosis
  • a baculovirus-dervived vector (see, Sarkis et al, Proc. Natl. Acad. Sci. US, Vol. 97(26): 14638-14643, 2000), encoding superoxide dismutase 1 (SODl), is added to a perfusion formulation such as:
  • This formulation is then perfused through a human central nervous system via ventriculo-lumbar perfusion for a period of 48 hours, thus transforming neurons to produce SODl.
  • adenoviruses adeno associated viruses, herpes viruses and lenti viruses are used to encode neuroprotective proteins calbindin, the anti-apoptotic proto-oncogene bcl-2 (see, e.g., Wei et al., J Neurochem 75(l):81-90, 2000), glial- derived neurotrophic growth factors (NGF, BDNF and NT3), epidermal growth factor (EGF), fibroblast growth factors (FGF 1-9), insulin (IGF-1), platelet-derived growth factors (PDGF-A, B &C), vascular endothelial growth factor (VEGF), and ciliary neurotrophic factor (CNTF).
  • NGF glial- derived neurotrophic growth factors
  • EGF epidermal growth factor
  • FGF 1-9 epidermal growth factor
  • IGF-1 insulin
  • PDGF-A, B &C platelet-derived growth factors
  • VEGF vascular endothelial growth factor
  • CNTF
  • An adenovirus vector expressing a cDNA encoding the protein leptin (see, Muzzin et al, Regulatory Peptides, 92(1-3 Special Issue SI): 57-64, 2000), is added to a perfusion formulation as follows:
  • This formulation is then perfused through a human central nervous system via ventriculo-lumbar perfusion for a period of 48 hours, thus transforming neurons to produce the satiety factor leptin.
  • antisense molecule is a molecule adapted to selectively bind to a nucleic acid in a cell to disrupt the target nucleic acid's function. These are typically
  • oligonucleotides but those of ordinary skill will recognize that the ordinary use of “antisense oligonucleotide” has come to refer to a variety of molecules based on standard nucleic acids but having various modifications and linkages, as now widely understood in the art. Thus, the term “antisense molecule” includes the ordinary meanings in the art of "antisense oligonucleotide.”
  • gene therapy includes any intervention in an animal (preferably a mammal, more preferably a human) that (i) causes a cell in the animal to express (as RNA or protein) a recombinant nucleic acid, whether such expression is transient or stable, (ii) causes a change in the cell's genome, such as an insertion, that changes the cell's pattern of gene expression, (iii) provides a transformed cell to the animal, or (iv) alters the expression of a gene in the cell.
  • gene therapy includes transformations with anti-sense constructs and uses of nucleic acid-based vaccines.
  • a nucleic acid for gene therapy can be an antisense molecule or a transforming nucleic acid, including a transforming nucleic acid adapted to direct the production of an antisense molecule.
  • a transforming nucleic acid can transform by positively encoding a gene product, inserting a promoter (such as in an appropriate gene trap such as described in U.S. Patent 6,080,576), disrupting a gene or gene expression, or the like.
  • • nutrient-providing effective amount is an amount that can be expected, provided sufficient amounts of other nutrients, to increase metabolism or reproduction of mammalian cells compared with nutrient solutions lacking that substance. • oncotic agent.
  • oncotic agent substances, generally macromolecules, that are of a size that is not readily able to leave the body cavity or other fluid containing body spaces (such as the cerebrospinal pathway, including the cerebral ventricles and subarachnoid spaces) into which they are inserted.
  • oncotic agents are exemplified by blood plasma expanders which are known in general as macromolecules having a size sufficient to inhibit their escape from the blood plasma through the circulatory capillary bed into the interstitial spaces of the body.
  • Serum albumin preferably human serum albumin, is one well known blood plasma protein that can be used as an oncotic agent.
  • Polysaccharide blood plasma expanders are often glucan polymers.
  • Hetastarch (a product of American Home Products) is an artificial colloid derived from a waxy starch composed almost entirely of amylopectin with hydroxyethyl ether groups introduced into the alpha (1-4) linked glucose units.
  • the colloid properties of a 6% solution (wt/wt) of hetastarch approximate those of human serum albumin.
  • Other polysaccharide derivatives may be suitable as oncotic agents in the blood substitute according to the invention. Among such other polysaccharide derivatives are hydroxymethyl alpha (1 -4) or (1-6) polymers and cyclodextrins. In general, it is preferred that the polysaccharide is one that is non-antigenic.
  • High molecular weight agents such as Dextran 70 having a molecular weight of about 70,000 Daltons are generally less preferred because they increase viscosity of the colloidal solution and impair the achievement of high flow rates.
  • the oncotic agent is in an amount effective to provide, in conjunction with other components of a fluorocarbon nutrient emulsion or a nutrient solution, an oncotic pressure of one to seven torr.
  • polynucleotide(s) or nucleic acid(s) generally refer to any polyribonucleotide or polydeoxyribonucleotide, which can be unmodified RNA or DNA or modified RNA or DNA.
  • Polynucleotide(s) include, without limitation, single- and double-stranded DNA, DNA that is a mixture of single- and double-stranded regions or single-, double- and triple-stranded regions, single- and double-stranded RNA, and RNA that is mixture of single- and double-stranded regions, hybrid molecules comprising DNA and RNA that can be single-stranded or, more typically, double-stranded, or triple-stranded regions, or a mixture of single- and double-stranded regions.
  • polynucleotide refers to triple-stranded regions comprising RNA or DNA or both RNA and DNA.
  • polynucleotide(s) also includes DNAs or RNAs as described above that contain one or more modified bases.
  • DNAs or RNAs with backbones modified for stability or for other reasons are “polynucleotide(s)” as that term is intended herein.
  • DNAs or RNAs comprising unusual bases, such as inosine, or modified bases, such as tritylated bases, to name just two examples are polynucleotides as the term is used herein.
  • polynucleotide(s) as it is employed herein embraces such chemically, enzymatically or metabolically modified fo ⁇ ns of polynucleotides, as well as the chemical forms of DNA and RNA characteristic of viruses and cells, including, for example, simple and complex cells, "Polynucleotide(s)” also embraces short polynucleotides often referred to as oligonucleotide(s).
  • respiration is the physical and chemical processes by which an organism supplies its cells and tissues with the oxygen needed for metabolism and, preferably, relieves them of the carbon dioxide formed in energy-producing reactions.
  • respiration-supporting amount is an amount that would, in model experiments, provide a statistically significant reduction in morbidity following a focal ischemic event.
  • transformed cell A cell is transformed if a recombinant nucleic acid is introduced into it or its ancestor so as to temporarily or stably (1) cause the cell to express a polypeptide or RNA in an amount not otherwise expressed by the cell or (2) interfere with the translation or transcription of a nucleic acid normally found in the cell.
  • a transforming composition is a composition containing a gene therapy effective amount of transformed cells or gene therapy agent.

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Abstract

L'invention porte sur un procédé d'application, sur un tissu neurologique, d'une composition de transformation ou d'une cellule transformée. Ce procédé consiste : (a) à injecter un liquide de perfusion cérébro-spinal physiologiquement acceptable dans un premier cathéter dans la voie cérébro-spinale, ce liquide perfusion cérébro-spinal possédant une quantité efficace en thérapie génique d'un agent de thérapie génique ou de cellules transformées ; (b) à prélever du liquide au moyen d'un deuxième cathéter et à l'injecter dans la voie cérébro-spinale afin de créer un flux et une voie de flux entre les premier et deuxième cathéters ; et (c) à conserver le flux pendant une durée suffisante pour perfuser au moins un volume de liquide céphalo-rachidien.
EP02713697A 2001-03-02 2002-02-28 Therapie genique pour tissus neurologiques Withdrawn EP1370138A4 (fr)

Applications Claiming Priority (3)

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US798775 1997-02-11
US79877501A 2001-03-02 2001-03-02
PCT/US2002/005886 WO2002069711A1 (fr) 2001-03-02 2002-02-28 Therapie genique pour tissus neurologiques

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EP1370138A1 true EP1370138A1 (fr) 2003-12-17
EP1370138A4 EP1370138A4 (fr) 2004-08-18

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WO2008115880A2 (fr) * 2007-03-21 2008-09-25 The Board Of Regents Of The University Of Texas System Utilisation de leptine pour le traitement ou la prévention de la maladie de parkinson

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WO2002005870A2 (fr) * 2000-07-19 2002-01-24 Neuron Therapeutics, Inc. Kits et compositions pour la realisation d'injections intracraniennes

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US5569180A (en) * 1991-02-14 1996-10-29 Wayne State University Method for delivering a gas-supersaturated fluid to a gas-depleted site and use thereof

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Publication number Priority date Publication date Assignee Title
WO2002005870A2 (fr) * 2000-07-19 2002-01-24 Neuron Therapeutics, Inc. Kits et compositions pour la realisation d'injections intracraniennes

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BACHOUD-LEVI A -C ET AL: "Neuroprotective gene therapy for Huntington's disease using a polymer encapsulated BHK cell line engineered to secrete human CNTF" HUMAN GENE THERAPY, vol. 11, no. 12, 10 August 2000 (2000-08-10), pages 1723-1729, XP002284732 ISSN: 1043-0342 *
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KORDOWER JEFFREY H ET AL: "Neurodegeneration prevented by lentiviral vector delivery of GDNF in primate models of Parkinson's disease" SCIENCE, AMERICAN ASSOCIATION FOR THE ADVANCEMENT OF SCIENCE,, US, vol. 290, no. 5492, 2000, pages 767-773, XP002207711 ISSN: 0036-8075 *
MCINNES RODERICK R ET AL: "Neural stem cells: A versatile tool for cell replacement and gene therapy in the central nervous system" CLINICAL GENETICS, vol. 56, no. 4, October 1999 (1999-10), pages 267-278, XP002284734 ISSN: 0009-9163 *
See also references of WO02069711A1 *

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WO2002069711A1 (fr) 2002-09-12

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