WO2008015023A2 - Utilisation de produits de photomédine pour la prévention et/ou le traitement de dysfonctionnements neuronaux - Google Patents

Utilisation de produits de photomédine pour la prévention et/ou le traitement de dysfonctionnements neuronaux Download PDF

Info

Publication number
WO2008015023A2
WO2008015023A2 PCT/EP2007/006933 EP2007006933W WO2008015023A2 WO 2008015023 A2 WO2008015023 A2 WO 2008015023A2 EP 2007006933 W EP2007006933 W EP 2007006933W WO 2008015023 A2 WO2008015023 A2 WO 2008015023A2
Authority
WO
WIPO (PCT)
Prior art keywords
photomedin
nucleic acid
product
acid molecule
human
Prior art date
Application number
PCT/EP2007/006933
Other languages
English (en)
Other versions
WO2008015023A3 (fr
Inventor
Daria Onichtchouk
Original Assignee
Develogen Aktiengesellschaft
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Application filed by Develogen Aktiengesellschaft filed Critical Develogen Aktiengesellschaft
Publication of WO2008015023A2 publication Critical patent/WO2008015023A2/fr
Publication of WO2008015023A3 publication Critical patent/WO2008015023A3/fr

Links

Classifications

    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K38/00—Medicinal preparations containing peptides
    • A61K38/16—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
    • A61K38/17—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans
    • A61K38/1703—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans from vertebrates
    • A61K38/1709—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans from vertebrates from mammals
    • 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
    • A61P25/14—Drugs for disorders of the nervous system for treating abnormal movements, e.g. chorea, dyskinesia
    • A61P25/16—Anti-Parkinson drugs
    • 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
    • A61P25/28—Drugs for disorders of the nervous system for treating neurodegenerative disorders of the central nervous system, e.g. nootropic agents, cognition enhancers, drugs for treating Alzheimer's disease or other forms of dementia
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K14/00—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
    • C07K14/435—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans
    • C07K14/46—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans from vertebrates
    • C07K14/47—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans from vertebrates from mammals

Definitions

  • the invention provides compositions and methods for treating and/or preventing neuronal dysfunctions, particularly neurodegenerative disorders.
  • the method of the invention involves administering to an individual in need of treatment a composition containing a Photomedin product and/or an effector/modulator thereof.
  • Parkinson's disease is a progressive neurodegenerative disorder; symptoms include uncontrolled tremors, stooped posture and gait disturbances. Morphologically, Parkinson's disease is characterized by a loss of the pigmented dopaminergic neurons located in the substantia nigra, resulting in depletion of the neurotransmitter dopamine. Other groups of neurons, such as the noradrenergic neurons of the locus coeruleus, can also be affected (reviewed by Zhang et al. Neurobiol Dis. 2000 7(4):240-250).
  • Parkinson's disease focus on replacement of dopamine, either through direct administration of its immediate precursor, levodopa, or by administration of dopamine receptor agonists or inhibitors of 15 dopamine metabolic enzymes, such as L- deprenyl (Grunblatt et al. J Neurol. 2000;247 Suppl 2:1195-1102).
  • dopamine receptor agonists or inhibitors of 15 dopamine metabolic enzymes such as L- deprenyl
  • multiple sclerosis This condition is a chronic, inflammatory CNS disease characterized pathologically by demyelination.
  • CNS disease characterized pathologically by demyelination.
  • RR-MS relapsing-remitting multiple sclerosis
  • SP- MS secondary progressive multiple sclerosis
  • PP-MS primary progressive multiple sclerosis
  • PR-MS progressive- relapsing multiple sclerosis
  • Photomedin-1 and Photomedin-2 Two novel olfactomedin-family extracellular proteins named Photomedin-1 and Photomedin-2. These proteins were secreted as disulphide-bonded dimers (Photomedin-1) or oligomers/multimers (Photomedin-2) with O-linked carbohydrate chains, although Photomedin-1 was proteolytically processed in the middle of the molecule after secretion. They found that Photomedin-1 was selectively expressed in the outer segment of photoreceptor cells in the retina and that Photomedin-2 was expressed in all retinal neurons. They also found that Photomedins preferentially bound to chondroitin sulphate-E and heparin. However, a function of Photomedins, in particular a function in vivo, has not been described.
  • WO 03/099318 describes the use of Photomedin products for preventing and/or treating pancreatic disorders, particularly those related to diabetes as well as neurodegenerative disorders.
  • WO 2005/051408 describes the use of Photomedin for stimulating and/or inducing the differentiation of development of insulin producing cells from progenitor cells.
  • Photomedins act as an survival factor for neuronal cells.
  • Photomedin products and effectors/molecules thereof are suitable as neuroprotective agents for inhibiting and/or preventing death of neuronal cells and/or for promoting survival and/or regeneration of neuronal cells.
  • Photomedin products are suitable for detecting and/or verifying or for the treatment, alleviation and/or prevention of neuronal dysfunctions such as neurodegenerative disorders and/or neuronal injury or trauma.
  • Photomedin products and effectors/modulators thereof have been identified as neuroprotective agents.
  • Neuroprotective agents 'Neuroprotection' refers to protection of the central or peripheral nervous system from neuronal loss, axonal loss and/or myelin loss.
  • Providing neuroprotection is one way to effect the prevention and/or treatment of neuronal dysfunctions such as neurodegenerative conditions and neurotrauma.
  • Zebrafish was used as model system for neurodegenerative disorders. Zebrafish has become a popular model vertebrate for the study of developmental processes as well as for pharmacological and toxicological studies over the last decade (Rubinstein A.L., (2003) Curr Opin Drug Discov Devel. 6: 218-223; Grunwald D.J. and Eisen J.S., (2002) Nat Rev Genet. 3: 717-724). In this organism, large numbers of transparent embryos which rapidly develop outside of their mother are readily available. Transgenic lines expressing fluorescent proteins under the control of tissue-specific promoters allow to rapidly assess the effects of pharmacological treatments or gene loss- and gain-of- function treatments.
  • zebrafish embryos represent a relevant model to identify genes or compounds which control neuronal cell formation in humans.
  • Zebrafish is known as established model system for human nervous system neurodegenerative diseases. Compounds that induce neurotoxicity in humans caused similar neurotoxicity in zebrafish. These results support the use of zebrafish as a predictive model for assessing neurotoxicity (Parng C, et al., J Pharmacol Toxicol Methods. 2006 May 6).
  • Transgenic zebrafish as models for neurodegenerative disorders have been described in WO 02/082043.
  • McKinley ET.et al Brain Res MoI Brain Res. 2005 Nov 30;141(2):128-137 show that the mechanism for dopaminergic neuron toxicity that is induced by a neurotoxin in mammals is proven to be conserved in zebrafish larvae.
  • the mechanism of spinal muscular atrophy (SMA) is similar in zebrafish and humans (Winkler et al., Genes Dev. 2005 Oct 1 ;19(19):2320-30).
  • zebrafish is used for studying the pathways underlying Alzheimer disease (Grilli et al., Funct Neurol. 2003 Jul-Sep;18(3):145-148; Tomasiewicz et al., J Neurosci Res. 2002 Dec 15;70(6):734-45)
  • the present invention relates to the use of a pharmaceutical composition
  • a pharmaceutical composition comprising a Photomedin product and/or an effector/modulator of said Photomedin product for the manufacture of a neuroprotective agent.
  • the composition contains pharmaceutically acceptable carriers, diluents and/or additives.
  • the pharmaceutical composition comprising the Photomedin product and/or the effector/modulator thereof may be used for diagnostic or therapeutic applications in vivo and/or in vitro. More particularly, the composition may be used as a neuroprotective agent for the treatment of neurodegenerative disorders, particularly neurodegenerative disorders of the central nervous system, e.g. involving the brain and/or the spinal cord, or of the peripheral nervous system.
  • the neurodegenerative disorder may be selected from Alzheimer's Disease, Parkinson's Disease, Amyotrophic Lateral Sclerosis (MS), Multiple Sclerosis or other diseases as indicated in more detail below.
  • the composition is suitable for the treatment of neuronal injury or trauma.
  • the composition is suitable for diagnostic purposes, e.g.
  • Photomedin-1 and Photomedin-2 are evolutionary conserved and present in the vertebrate organisms from zebrafish to human (see Fig. 5).
  • Photomedin-1 and Photomedin-2 have four homologues (zPhotomedin-1a, zPhotomedin-1 b, zPhotomedin-2a, and zPhotomedin-2b).
  • Photomedin product includes Photomedin homologous proteins and nucleic acid molecules coding therefor, which are obtainable from vertebrate, e.g. fish, avian or mammalian species. Particularly preferred are nucleic acids encoding the human Photomedin protein and variants thereof.
  • the invention particularly relates to a nucleic acid molecule encoding a polypeptide contributing to neuroprotection, wherein said nucleic acid molecule comprises
  • (f) a partial sequence of any of the nucleotide sequences of (a) to (e) having a length of 15-25 bases, preferably 25-35 bases, more preferably 35-50 bases and most preferably at least 50 bases.
  • Photomedin proteins include purified natural, synthetic, or recombinant Photomedin-1 or Photomedin-2 and variants thereof. Variants include insertion, substitution and deletion variants and chemically modified derivatives. Variants also include recombinant proteins, for example but not limited to hybrids or fusions of Photomedin-1 or Photomedin-2 and other proteins. Also included are proteins or peptides substantially homologous to the human Photomedin-1 or Photomedin-2 protein having the amino acid sequence published as GenBank Accession Number XP_034000 (Photomedin-1 ) or GenBank Accession Number NP_872293 (Photomedin- 2). The term "Photomedin product" also includes nucleic acids, e.g.
  • Photomedin product also includes Photomedin-1 or Photomedin-2 homodimers or heterodimers of a Photomedin-1 or Photomedin-2 protein product and another protein.
  • the present invention also refers to proteins which have a high degree of identity to the biologically active Photomedin protein product having the amino acid sequence published as GenBank Accession Number XP_034000 (Photomedin-1) or GenBank Accession Number NP_872293 (Photomedin-2), that is preferably in excess of 70%, most preferably in excess of 80%, and even more preferably in excess of 90% or 95% over the entire protein.
  • the degree of identity between the mouse and the human protein is about 91%, and it is contemplated that preferred mammalian Photomedin proteins will have a similarly high degree of identity.
  • the percentage of homology or percent identity between a Photomedin product and a human Photomedin protein or nucleic acid may be determined according to standard procedures, e.g. by using the BLAST algorithm. Preferably, it is calculated as the percentage of nucleotide or amino acid residues found in the smaller of the two sequences that align with identical nucleotide or amino acid residues in the sequence being compared, when four gaps in a length of 100 nucleotides or amino acids may be introduced to assist in that alignment.
  • substantially homologous is any Photomedin protein product which may be isolated by virtue of cross-reactivity with antibodies to the Photomedin protein product or whose genes may be isolated through hybridization with the gene or with segments of the gene encoding the Photomedin protein product.
  • effector/modulator of a Photomedin product refers to any substance that modulates, induces or e.g. stimulates or inhibits the expression and/or function of Photomedin.
  • E ⁇ xamples of effectors/modulators of Photomedin products particularly refer to biologically active nucleic acids such as antisense molecules, ribozymes, siRNA molecules or precursors thereof.
  • the term encompasses antibodies or antibody fragments, aptamers, or scaffold proteins specifically directed against a Photomedin protein product.
  • effector/modulator also refers to hybridization probes and/or primers.
  • the neurodegenerative disease may be selected from Alzheimer disease, Parkinson disease, Creutzfeldt-Jakob disease, Huntington disease, Multiple sclerosis, Alper's disease, Amyotrophic lateral sclerosis, Ataxia telangiectasia, Alexander disease, Batten disease (also known as Spielmeyer-Vogt-Sjogren-Batten disease, BSE, Canavan disease, Cockayne syndrome, Corticobasal degeneration, Kennedy's disease, Krabbe disease, Lewy body dementia, Machado-Joseph disease (Spinocerebellar ataxia type 3), Multiple System Atrophy, Olivopontocerebellar Atrophy, Pelizaeus-Merzbacher Disease, Progressive Supranuclear Palsy, Pick's disease, Postpolyomielitus syndrome, Primary lateral sclerosis, Refsum's disease, Sandhoff disease, Schilder's disease, Shy-Drager Syndrome, Spielmeyer-Vogt-Sjogren
  • the medicament comprises a Photomedin protein product as an active ingredient.
  • the Photomedin protein product may be a native or recombinant protein including fusion hybrid proteins.
  • the protein may be a glycosylated or non-glycosylated protein.
  • the heterologous signal sequence selected should be one that is recognized and processed, i.e., cleaved by a signal peptidase, by the host cell.
  • the signal sequence may be substituted by a prokaryotic signal sequence selected, for example, from the group of the alkaline phosphatase, penicillinase, or heat-stable enterotoxin 11 leaders.
  • the native Photomedin-1 or Photomedin-2 signal sequence may be substituted by the yeast invertase, alpha factor, or acid phosphatase leaders.
  • the native signal sequence is satisfactory, although other mammalian signal sequences may be suitable.
  • Expression and cloning vectors generally include a nucleic acid sequence that enables the vector to replicate in one or more selected host cells.
  • polynucleotide sequences that are capable of hybridizing to the claimed nucleotide sequences, and in particular, those of the polynucleotide encoding the proteins of the invention, under various conditions of stringency.
  • Hybridization conditions are based on the melting temperature (Tm) of the nucleic acid binding complex or probe, as described in Wahl G.M. et al., (1987; Methods Enzymol. 152: 399-407) and Kimmel A.R. (1987; Methods Enzymol. 152: 507-511), and may be used at a defined stringency.
  • hybridization under stringent conditions means that after washing for 1 h with 1 x SSC and 0.1% SDS at 50 0 C, preferably at 55°C, more preferably at 62°C and most preferably at 65°C, particularly for 1 h in 0.2 x SSC and 0.1% SDS at 50 0 C, preferably at 55 0 C, more preferably at 62 0 C and most preferably at 65°C, a positive hybridization signal is observed.
  • Altered nucleic acid sequences encoding the proteins which are encompassed by the invention include deletions, insertions or substitutions of different nucleotides resulting in a polynucleotide that encodes the same or a functionally equivalent protein.
  • the encoded proteins may also contain deletions, insertions or substitutions of amino acid residues, which produce a silent change and result in functionally equivalent proteins. Deliberate amino acid substitutions may be made on the basis of similarity in polarity, charge, solubility, hydrophobicity, hydrophilicity, and/or the amphipathic nature of the residues as long as the biological activity of the protein is retained.
  • Photomedin fragments e.g. peptide fragments of the proteins or derivatives thereof such as cyclic peptides, retro-inverso peptides or peptide mimetics having a length of at least 4, preferably at least 6 and up to 300 amino acids.
  • the fragments may contain at least one Photomedin CXCXCX(9) C motif, where C is cysteine and X is any amino acid.
  • the CXCXCX(9)C motive is specific for N-termini of mature Photomedins 1 and 2 and conserved across species. It is involved in cysteine bond formation and dimerisation of Photomedins.
  • these fragments comprise amino acids 21-260 of human Photomedin-2 or Photomedin-1 or at least 100 contiguous amino acids thereof or a sequence having an identity of at least
  • peptides could be fragments encompassing the olfactomedin domain of Photomedin-2 or Photomedin-1 , or parts of it.
  • these fragments comprise amino acids 494-744 of mouse Photomedin-2 (olfactomedin-like 2B, Accession Number NP_796042); amino acids 498-748 of human Photomedin-2 (olfactomedin- like 2B, GenBank Accession Number NP_056256); amino acids 445-679 of mouse Photomedin-1 (olfactomedin-like 2A, GenBank Accession Number NP_766442), or amino acids 202-436 of human Photomedin-1 (Olfactomedin-like 2A GenBank Accession Number AAH54001 ) or at least 100 contiguous amino acids thereof or a sequence having an identity of at least 85%, preferably at least 90% and more preferably at least 95% thereto.
  • allelic variants of the genes encoding the proteins of the invention are also included within the scope of the present invention.
  • an 'allele 1 or 'allelic sequence' is an alternative form of the gene, which may result from at least one mutation in the nucleic acid sequence. Alleles may result in altered mRNAs or polypeptides whose structures or function may or may not be altered. Any given gene may have none, one or many allelic forms.
  • Common mutational changes, which give rise to alleles are generally ascribed to natural deletions, additions or substitutions of nucleotides. Each of these types of changes may occur alone or in combination with the others, one or more times in a given sequence.
  • the nucleotide sequences encoding the proteins may be inserted into appropriate expression vectors, i.e., a vector which contains the necessary elements for the transcription and translation of the inserted coding sequence.
  • appropriate expression vectors i.e., a vector which contains the necessary elements for the transcription and translation of the inserted coding sequence.
  • Methods which are well known to those skilled in the art, may be used to construct expression vectors containing sequences encoding the proteins and the appropriate transcriptional and translational control elements.
  • Regulatory elements include for example a promoter, an initiation codon, a stop codon, a mRNA stability regulatory element, and a polyadenylation signal.
  • a polynucleotide can be assured by (i) constitutive promoters such as the Cytomegalovirus (CMV) promoter/enhancer region, (ii) tissue specific promoters such as the insulin promoter (see, Soria B. et al., (2000), Diabetes 49: 157-162), SOX2 gene promoter (see Li M. et al., (1998) Curr. Biol. 8: 971-974), Msi-1 promoter (see Sakakibara S. and Okano H., (1997) J. Neuroscience 17: 8300-8312), alpha-cardia myosin heavy chain promoter or human atrial natriuretic factor promoter (Klug M.G.
  • constitutive promoters such as the Cytomegalovirus (CMV) promoter/enhancer region
  • tissue specific promoters such as the insulin promoter (see, Soria B. et al., (2000), Diabetes 49: 157-162), SOX2 gene promoter (
  • natural, modified or recombinant nucleic acid sequences encoding the proteins of the invention and homologous proteins may be ligated to a heterologous sequence to encode a fusion protein.
  • Heterologous sequences are preferably located at the N- and/or C-terminus of the fusion protein.
  • Means for producing labeled hybridization or PCR probes for detecting polynucleotide sequences include oligo-labeling, nick translation, end-labeling of RNA probes, PCR amplification using a labeled nucleotide, or enzymatic synthesis. These procedures may be conducted using a variety of commercially available kits (Pharmacia & Upjohn, (Kalamazoo, Mich.); Promega (Madison Wis.); and U.S. Biochemical Corp., (Cleveland, Ohio).
  • Suitable reporter molecules or labels include radionuclides, enzymes, fluorescent, chemiluminescent or chromogenic agents as well as substrates, co-factors, inhibitors, magnetic particles, and the like.
  • Specific constructs of interest include anti-sense molecules, which will block the expression of the proteins of the invention, or expression of dominant negative mutations.
  • a detectable marker such as for example lac-Z, may be introduced in the locus of the genes of the invention, where up-regulation of expression of the genes of the invention will result in an easily detected change in phenotype.
  • One may also provide for expression of the genes of the invention or variants thereof in cells or tissues where it is not normally expressed or at abnormal times of development.
  • by providing expression of the proteins of the invention in cells in which they are not normally produced one can induce changes in cell behavior.
  • nucleic acids and proteins of the invention are, for example but not limited to, the following: (i) protection of nervous system (neuroprotection and protection of all cell types comprising nervous system and cell types interacting with the nervous system ) in vitro and in vivo, (ii) nervous tissue regeneration (regeneration of nervous system and regeneration of all cell types composing nervous system and cell types interacting with the nervous system) in vitro and in vivo, (iii) small molecule drug target, (iv) antibody target (therapeutic, diagnostic, drug targeting/cytotoxic antibody), (v) diagnostic and/or prognostic marker, (vi) protein therapy, (vii) gene therapy (gene delivery/gene ablation), and / or (viii) research tools.
  • the polynucleotides and the proteins of the invention and particularly the human polynucleotides and polypeptides may be useful in preventing or regulating the degeneration of tissues or preventing the cell death of tissues when administered to a subject in need thereof.
  • the compositions of the present invention will have efficacy for treatment of patients suffering from neurodegenerative disorders.
  • Photomedin nucleic acids and proteins and effectors/modulators thereof are useful in diagnostic and therapeutic applications implicated in various embodiments as described below.
  • Photomedin polynucleotides may be useful in gene therapy, and the Photomedin proteins may be useful when administered to a subject in need thereof.
  • the compositions of the present invention will have efficacy for treatment of patients suffering from neuronal dysfunctions, e.g. neurodegenerative disorders as described above.
  • the Photomedin nucleic acids and proteins and effectors/modulators thereof may be administered either as a monotherapy or as a combination therapy with other pharmaceutical agents.
  • they may be administered together with other pharmaceutical agents suitable for the treatment and/or prevention of neuronal dysfunction, e.g. neurodegenerative disorders.
  • they may be administered together with pharmaceutical agents which have an immunosuppressive activity, e.g. antibodies, polypeptides and/or peptidic or non-peptidic low molecular weight substances.
  • immunosuppressive agents are listed in the following Table 1.
  • Table 1 Exemplary agents for immune suppression
  • the combination therapy may comprise coadministration of the medicaments during the treatment period and/or separate administration of single medicaments during different time intervals in the treatment period.
  • compositions may comprise Photomedin nucleic acids and/or the proteins and/or effectors/modulators thereof such as antibodies, mimetics, agonists, antagonists or inhibitors of the proteins or nucleic acids as an active ingredient.
  • the compositions may be administered alone or in combination with at least one other agent, such as stabilizing compound, which may be administered in any sterile, biocompatible pharmaceutical carrier, including, but not limited to, saline, buffered saline, dextrose, and water.
  • the compositions may be administered to a patient alone or in combination with other agents, drugs or hormones.
  • compositions utilized in this invention may be administered by any number of routes including, but not limited to, oral, intravenous, intramuscular, intra-arterial, intramedullary, intrathecal, intraventricular, transdermal, subcutaneous, intraperitoneal, intranasal, enteral, topical, sublingual or rectal means.
  • routes including, but not limited to, oral, intravenous, intramuscular, intra-arterial, intramedullary, intrathecal, intraventricular, transdermal, subcutaneous, intraperitoneal, intranasal, enteral, topical, sublingual or rectal means.
  • the composition is administered by injection, e.g. by subcutaneous, intravenous or intramuscular injection.
  • these pharmaceutical compositions may contain suitable pharmaceutically-acceptable carriers comprising excipients and auxiliaries, which facilitate processing of the active compounds into preparations, which can be used pharmaceutically. Further details on techniques for formulation and administration may be found in the latest edition of Remington's Pharmaceutical Sciences (Maack Publishing Co., Easton, Pa.).
  • compositions suitable for use in the invention include compositions wherein the active ingredients are contained in an effective amount to achieve the intended purpose.
  • the determination of an effective dose is well within the capability of those skilled in the art.
  • the therapeutically effective dose can be estimated initially either in cell culture assays, e.g., of preadipocyte cell lines or in animal models, usually mice, rabbits, dogs or pigs.
  • the animal model may also be used to determine the appropriate concentration range and route of administration. Such information can then be used to determine useful doses and routes for administration in humans.
  • a therapeutically effective dose refers to that amount of active ingredient, for example the Photomedin nucleic acids or proteins or fragments thereof or antibodies, which is sufficient for treating a specific condition.
  • Therapeutic efficacy and toxicity may be determined by standard pharmaceutical procedures in cell cultures or experimental animals, e.g., ED50 (the dose therapeutically effective in 50% of the population) and LD50 (the dose lethal to 50% of the population).
  • the dose ratio between therapeutic and toxic effects is the therapeutic index, and it can be expressed as the ratio, LD50/ED50.
  • Pharmaceutical compositions, which exhibit large therapeutic indices, are preferred. The data obtained from cell culture assays and animal studies is used in formulating a range of dosage for human use.
  • Long-acting pharmaceutical compositions may be administered every 3 to 4 days, every week or once every two weeks depending on half-life and clearance rate of the particular formulation.
  • Normal dosage amounts may vary from 0.01 to 100,000 ⁇ g, up to a total dose of about 1 g, depending upon the route of administration.
  • An especially preferred dosage is from 70 ng to 1 ⁇ g per kg body weight per injection.
  • Guidance as to particular dosages and methods of delivery is provided in the literature and generally available to practitioners in the art. Those skilled in the art employ different formulations for nucleotides than for proteins or their inhibitors. Similarly, delivery of polynucleotides or polypeptides will be specific to particular cells, conditions, locations, etc.
  • Fig. 1 shows expression of Photomedin-2 homologues in developing zebrafish embryo (in-situ hybridisation) (shown in purple). A first expression can be noticed at 24 hours post fertilisation (hpf). Top panel: Photomedin-2a is expressed as a stripe aligning otic vesicle (ear primordia). Bottom panel: Photomedin-2b is expressed in the boundary between Midbrain and Hindbrain (MHB).
  • Fig. 2 shows expression of Photomedin-1 homologues in zebrafish (in-situ hybridisation) (shown in purple).
  • Fig. 2a shows expression of Photomedin-1 a at indicated developmental stages.
  • side view shows Photomedin-1 b is expressed in the deep layers in the dorsal midline (top left).
  • Top (top right) and side (bottom left) of 17 hpf embryo views show expression in the nervous system ganglia.
  • polynucleotide comprising the nucleotide sequence as shown in GenBank Accession number relates to the expressible gene of the nucleotide sequences deposited under the corresponding GenBank Accession number.
  • GenBank Accession number relates to NCBI GenBank database entries (Ref.: Benson D.A. et al., (2000) Nucleic Acids Res. 28: 15-18).
  • Photomedin proteins and nucleic acid molecules coding therefore are obtainable from vertebrate species, e.g. mammals or fish. Particularly preferred are nucleic acid molecules and proteins encoced thereby comprising human and mouse.
  • NCBI non redundant protein database [ftp://ftp.ncbi.nih.gov/blast/db]
  • EST section of NCBI Genbank see Boguski et al., 1993, Nat Genet. 4: 332-333.
  • dbEST-database for "expressed sequence tags” and zebrafish genome draft assembly 2 [http://www.ensembl.org/Danio rerio/1) were searched using the blastall programm (version 2.2.6, Altschul et al. 1997, Nucleic Acids Res. 25: 3389- 3402).
  • Zebrafish were raised, maintained, and crossed as described (see, Westerfield M., (1995) The Zebrafish Book. Eugene, Oregon: Univ.of Oregon Press). Staging was performed according to Kimmel CB. et al., (1995) Dev Dyn 203: 253-310. Development of zebrafish embryos was carried out at 28°C. The age of embryos is indicated as hours post fertilization (hpf).
  • antisense morpholino oligonucleotides were used. If available from the assembly data (see Example 1 and Fig. 5), translation start sites were selected for antisense oligonucleotide targeting. Otherwise splice donor sites identified by alignment of zebrafish EST data or mouse protein data to zebrafish genomic sequence were used for antisense oligonucleotides targeting. The following morpholino oligonucleotides were used: PH2a: 2 splice donor blockers
  • SEQ ID NO:9 S'-AATGTATCTTACCTGACTTAATATA-a' SEQ ID NO:10 ⁇ '-CTGGTCACCTTGATATTATCTTTTC-S' PH2b 1 translation blocker
  • SEQ ID NO: 12 5'-GCTCCACAATCCTCCACATCTCGGC-3' PH1b 2 splice donor blockers
  • SEQ ID NO:13 5'-TTACCTCTTCTAATGTGTTCATCTG-3'
  • SEQ ID NO: 14 5 I -TGTGTTACCTGTGCTTTACTGTCTG-3 I
  • Photomedin or control antisense oligonucleotides were injected into fertilized one-cell stage embryos as described (see, for example, Nasevicius & Ekker, 2000, Nat Genet 26: 216-220; Urtishak et al., 2003, Dev Dyn. 228: 405-413). Injected embryos were analysed at different stages of development.(Fig.3 and Fig 4). We found in this invention that 'translation blocker' morpholino blocks the expression of maternal as well as zygotic RNA whereas the 'splice blocker' morpholino blocks only zygotic one.
  • the loss-of function phenotype of Photomedin-1a (PH 1a) is significantly more severe than the main loss-of-function phenotype of Photomedin-1b (PH1 b), although the pattern of expression of these genes is quite similar (see Fig.2).
  • Example 4 Acridine orange stain of living zebrafish for identification of apoptotic cells.
  • Photomedin-2 and Photomedin-1 contain a signal peptide followed by N- terminal cystein-rich domain, separated by a spacer region from C-terminal olfactomedin domain.
  • Photomedin-2 contains two potential N-glycosylation sites at amino acid residues 187 and 213 (Fig.6 a), top).
  • the N-terminal domain contains two cystein stretches CXCXCX(9)C, conserved between homologues in vertebrates (including zebrafish, mouse, rat and human), suggesting N-terminal disulfide-bonded dimer formation (see Figure 5).

Landscapes

  • Health & Medical Sciences (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Chemical & Material Sciences (AREA)
  • Engineering & Computer Science (AREA)
  • Bioinformatics & Cheminformatics (AREA)
  • General Health & Medical Sciences (AREA)
  • Biomedical Technology (AREA)
  • Neurology (AREA)
  • Neurosurgery (AREA)
  • Organic Chemistry (AREA)
  • Medicinal Chemistry (AREA)
  • Veterinary Medicine (AREA)
  • Public Health (AREA)
  • Pharmacology & Pharmacy (AREA)
  • Animal Behavior & Ethology (AREA)
  • Chemical Kinetics & Catalysis (AREA)
  • Gastroenterology & Hepatology (AREA)
  • General Chemical & Material Sciences (AREA)
  • Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
  • Proteomics, Peptides & Aminoacids (AREA)
  • Zoology (AREA)
  • Epidemiology (AREA)
  • Genetics & Genomics (AREA)
  • Marine Sciences & Fisheries (AREA)
  • Psychology (AREA)
  • Toxicology (AREA)
  • Biochemistry (AREA)
  • Biophysics (AREA)
  • Immunology (AREA)
  • Molecular Biology (AREA)
  • Hospice & Palliative Care (AREA)
  • Psychiatry (AREA)
  • Medicines That Contain Protein Lipid Enzymes And Other Medicines (AREA)
  • Acyclic And Carbocyclic Compounds In Medicinal Compositions (AREA)
  • Pharmaceuticals Containing Other Organic And Inorganic Compounds (AREA)

Abstract

L'invention concerne des compositions et des procédés destinés au traitement et/ou à la prévention de dysfonctionnements neuronaux, en particulier des maladies neurodégénératives. Le procédé de l'invention concerne l'administration, à un individu nécessitant un traitement, d'une composition contenant un produit de photomédine et/ou un effecteur/modulateur de celui-ci.
PCT/EP2007/006933 2006-08-04 2007-08-06 Utilisation de produits de photomédine pour la prévention et/ou le traitement de dysfonctionnements neuronaux WO2008015023A2 (fr)

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
EP06016326.8 2006-08-04
EP06016326 2006-08-04

Publications (2)

Publication Number Publication Date
WO2008015023A2 true WO2008015023A2 (fr) 2008-02-07
WO2008015023A3 WO2008015023A3 (fr) 2008-05-22

Family

ID=38922813

Family Applications (1)

Application Number Title Priority Date Filing Date
PCT/EP2007/006933 WO2008015023A2 (fr) 2006-08-04 2007-08-06 Utilisation de produits de photomédine pour la prévention et/ou le traitement de dysfonctionnements neuronaux

Country Status (1)

Country Link
WO (1) WO2008015023A2 (fr)

Citations (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2003099318A2 (fr) * 2002-05-29 2003-12-04 DeveloGen Aktiengesellschaft für entwicklungsbiologische Forschung Proteines specifiques du pancreas
WO2005051408A1 (fr) * 2003-11-28 2005-06-09 Develogen Aktiengesellschaft Procede de prevention et traitement du diabete par la dg119

Patent Citations (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2003099318A2 (fr) * 2002-05-29 2003-12-04 DeveloGen Aktiengesellschaft für entwicklungsbiologische Forschung Proteines specifiques du pancreas
WO2005051408A1 (fr) * 2003-11-28 2005-06-09 Develogen Aktiengesellschaft Procede de prevention et traitement du diabete par la dg119

Non-Patent Citations (1)

* Cited by examiner, † Cited by third party
Title
FURUTANI YUTAKA ET AL: "Identification and characterization of photomedins: novel olfactomedin-domain-containing proteins with chondroitin sulphate-E-binding activity." THE BIOCHEMICAL JOURNAL 1 AUG 2005, vol. 389, no. Pt 3, 1 August 2005 (2005-08-01), pages 675-684, XP002465216 ISSN: 1470-8728 cited in the application *

Also Published As

Publication number Publication date
WO2008015023A3 (fr) 2008-05-22

Similar Documents

Publication Publication Date Title
US7425334B2 (en) Methods of making antibodies that bind Nogo
KR100743376B1 (ko) 신경영양성 인자
US6777388B1 (en) Leptin-related peptides
US9644027B2 (en) Antibodies that specifically bind to DRG11-responsive (DRAGON) proteins
US20030215913A1 (en) Nucleic acids, vectors, host cells, polypeptides, and uses thereof
KR20100017494A (ko) 프로테오글리칸 폴리펩타이드를 암호화하는 핵산을 이용한 상처 및 피부손상의 치료
CN1372569A (zh) 角质形成细胞生长因子－2
KR100640265B1 (ko) Ｌｙ6ｈ 유전자
US9988427B2 (en) Erythropoietin variants
EP2096120A2 (fr) Utilisation de produits à protéines sécrétées pour empêcher et traiter les maladies pancréatiques et/ou l'obésité et/ou un syndrome métabolique
KR20040089093A (ko) 시스틴-결절 폴드 단백질
WO2008015023A2 (fr) Utilisation de produits de photomédine pour la prévention et/ou le traitement de dysfonctionnements neuronaux
EP2269628A2 (fr) Proteines spécifiques du pancréas
EP1450852B1 (fr) Acides nucleiques et peptides de ptp10d intervenant dans la regulation de l'homeostasie energetique
US20030211991A1 (en) Human sez6 nucleic acids and polypeptides
WO2002002771A2 (fr) Nouveau gene hsparc-11 et procedes de fabrication et d'utilisation de ce dernier
DE60024862T2 (de) "insulinabhängiges sequenz dna bindendes protein-1" (irsdbp-1), dafür kodierendes gen und ihre verwendungen
EP1290026A2 (fr) Acides nucleiques et polypeptides sez6 humains
WO2004020465A2 (fr) Proteines intervenant dans la regulation de l'homeostasie energetique
WO2004012758A1 (fr) Utilisation de tgf beta ig-h3 dans la prevention et le traitement de l'obesite, du diabete et/ou du syndrome metabolique
WO2002059369A2 (fr) Compositions et procedes de traitement des maladies liees a une mauvaise regulation du cholesterol
WO2001066738A2 (fr) Acides nucleiques codant des polypeptides crsp1 humains et utilisations de ces derniers
US20040197900A1 (en) Foggy
WO2005067965A2 (fr) Utilisation de produits proteiques permettant de prevenir et de traiter les maladies du pancreas et/ou l'obesite et/ou le syndrome metabolique
US20040215007A1 (en) Novel secreted proteins and their uses

Legal Events

Date Code Title Description
121 Ep: the epo has been informed by wipo that ep was designated in this application

Ref document number: 07786578

Country of ref document: EP

Kind code of ref document: A2

NENP Non-entry into the national phase in:

Ref country code: DE

NENP Non-entry into the national phase in:

Ref country code: RU

122 Ep: pct application non-entry in european phase

Ref document number: 07786578

Country of ref document: EP

Kind code of ref document: A2