WO2007113366A1 - Utilisation de composés induisant le développement axonal des neurones, compositions thérapeutiques contenant ces composés et leurs applications - Google Patents

Utilisation de composés induisant le développement axonal des neurones, compositions thérapeutiques contenant ces composés et leurs applications Download PDF

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WO2007113366A1
WO2007113366A1 PCT/ES2007/070068 ES2007070068W WO2007113366A1 WO 2007113366 A1 WO2007113366 A1 WO 2007113366A1 ES 2007070068 W ES2007070068 W ES 2007070068W WO 2007113366 A1 WO2007113366 A1 WO 2007113366A1
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seq
nrgl
human
sequence
amino acid
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PCT/ES2007/070068
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Spanish (es)
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Oscar MARÍN PARRA
Beatriz Rico Gozalo
Guillermina LÓPEZ BENDITO
Nuria Flames Bonilla
Sonia Lorezno Vidal
Juan Antonio SÁNCHEZ ALCAÑIZ
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Consejo Superior De Investigaciones Científicas
Universidad Miguel Hernández De Elche
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Publication of WO2007113366A1 publication Critical patent/WO2007113366A1/fr

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    • 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
    • A61K38/18—Growth factors; Growth regulators
    • A61K38/1883—Neuregulins, e.g.. p185erbB2 ligands, glial growth factor, heregulin, ARIA, neu differentiation factor

Definitions

  • Biomedicine and biotechnology Development of active therapeutic principles. Development of pharmaceutical compositions for the treatment of human diseases that occur with alterations of the axonal development of neurons.
  • axonal growth is very restricted from the last stages of its development. For this reason, the regeneration of axons after trauma or neurodegenerative diseases is one of the most important challenges of current biomedicine.
  • the restriction of axonal growth in the CNS is mainly due to the existence of factors that slow axonal growth in the adult brain and the absence of factors that promote axon elongation.
  • factors that restrict the growth of axons in recent years much progress has been made in the study of several proteins derived from myelin, such as Nogo, MAG and OMgp.
  • the regulation of axonal growth, the formation of collaterals and new connections in the postnatal brain by myelin proteins is a research area with great therapeutic potential.
  • axons Among the factors that promote the extension of axons, one of the recently used strategies and that constitutes a very active field of research is the use of enveloping glia. Other factors that promote the growth of Certain axons are Netrin-1 and brain-derived growth factor (BDNF).
  • BDNF brain-derived growth factor
  • Axonal regeneration is frequently seen from the perspective of spinal injuries, but there are numerous neuronal pathologies in which the axons of other populations of CNS neurons are affected, such as in schizophrenia, Alzheimer's disease or multiple sclerosis (Principies of Neural Science (2000) Kandel ER, Schwartz JH, Jessell TM Eds. McGraw-Hill, New York).
  • schizophrenia for example, there is a deficit of projections from axons of the mediodorsal thalamus
  • Interneurons are characterized by having short axons, unlike neurons with long axons such as cortical-spinal or thalamus-cortical, but in any case their regeneration could represent an improvement considerable in cortical functioning and therefore represents an attractive target for the development of an appropriate therapy for this disease.
  • NRG Neuregulins
  • NRGl In the nervous system, NRGl is essential for the survival of glia cells, as well as for their proliferation and differentiation (Buonanno A, Fischbach GD (2001) Neuregulin and ErbB receptor signaling pathways in the nervous system. Curr Opin Neurobiol 11: 287- 296). In neurons, NRG1 functions range from proliferation control to very late events in neuronal development such as the regulation of channel expression (Buonanno A, Fischbach GD (2001) Neuregulin and ErbB receptor signaling pathways in the nervous system Curr Opin Neurobiol 11: 287-296).
  • NRGl is also able to promote the migration of Schwann cells and cortical neurons (Mahanthappa NK, Anton ES, Matthew WD (1996) Glial growth factor 2, to soluble neuregulin, directly increases Schwann cell motility and indirectly promotes neurite outgrowth. J Neurosci 16: 4673-4683; Flames N, Long JE, Garratt AN, Fischer TM, Gassmann M, Birchmeier C, Lai C, Rubenstein JL, Mar ⁇ n 0 (2004) Short- and long-range attraction of cortical GABAergic interneurons by neuregulin-1.
  • Neuron 44: 251-261 to act as a memory modulator (Use of neuregulin-beta as an indicator and / or target, US Patent 20030036101), to produce mammalian glial cells (Methods for differentiating neural stem cells to glial cells using neuregulins, US patent 6,033,906) and useful in the treatment of schizophrenia (Human schizophrenia gene, US Patent 20050208527).
  • neuregulins are therapeutic compounds for different human diseases and can be used for others not yet identified.
  • An object of the present invention is the use of a compound that induces the activity of neuregulin 1 (hereinafter, NRGl), hereinafter used an activating compound of the present invention, for the preparation of a medicament or pharmaceutical composition for the treatment of a disease caused by a impaired axonal development of neurons of the nervous system.
  • NRGl neuregulin 1
  • a particular object of the invention is the use of a compound that induces NRG1 activity based on the use of a nucleotide sequence, hereinafter NRG1 nucleotide sequence of the present invention, which allows the expression of a protein or peptide inducing the axonal regeneration of human neurons, and which is constituted by one or several nucleotide sequences belonging to the following group: a) a nucleotide sequence consisting of the nucleotide sequence encoding human NRGl ⁇ (SEQ ID NOl), b) a nucleotide sequence consisting of the nucleotide sequence encoding human NRGlCC (SEQ ID N03), c) a nucleotide sequence analogous to the sequence of a) and b), d) a fragment of any one of the sequences of a), b) and c), and e) a nucleotide sequence, genetic construct, comprising any sequence belonging to a), b), c) and
  • another particular object of the invention is the use of an inducing compound of the invention in which the activating molecule is a protein or peptide, hereinafter use of the NRG1 protein of the present invention, which exhibits axon-developing activity of human neurons, and comprising one or more amino acid sequences belonging to the following group: a) an amino acid sequence consisting of the human NRGl ⁇ amino acid sequence (SEQ ID N02), b) an amino acid sequence consisting of the human NRGlOC amino acid sequence (SEQ ID N02), b) an amino acid sequence consisting of the human NRGlOC amino acid sequence (SEQ ID N02), b) an amino acid sequence consisting of the human NRGlOC amino acid sequence (SEQ ID N02), b) an amino acid sequence consisting of the human NRGlOC amino acid sequence (SEQ ID N02), b) an amino acid sequence consisting of the human NRGlOC amino acid sequence (SEQ ID N02), b) an amino acid sequence consisting of the human NRGlOC amino acid
  • NRG1 cells of the invention are preferably human eukaryotes and more preferably neurons- or prokaryotes, hereinafter NRG1 cells of the invention, genetically modified and comprising the nucleotide sequence, the construction or the vector of NRG1 expression of the invention -in which the NRG peptide or protein of the invention can be adequately expressed- for the preparation of an inducing compound of the invention and its subsequent purification and application.
  • Another object of the present invention is a pharmaceutical composition or medicament for the treatment of diseases, disorders or pathologies that occur with alterations in the development of neuron axons, hereinafter pharmaceutical composition of the present invention, comprising a compound or inducing agent of the invention, in therapeutically effective amount together with, optionally, one or more pharmaceutically acceptable adjuvants and / or vehicles.
  • a particular embodiment of the invention is a pharmaceutical composition of the invention in which the compound or agent capable of developing neuron axons is one or several NRG1 nucleotide sequences belonging to the following group: a) a nucleotide sequence consisting of the sequence nucleotide encoding the NRGl ⁇ (SEQ ID NOl), b) a nucleotide sequence consisting of the nucleotide sequence encoding the NRGlCC
  • Another particular object of the present invention is a pharmaceutical composition of the invention in which the compound or agent capable of recovering the development of neuron axons is a protein or a peptide encoded by the sequence, genetic construct or NRGl vector of the invention.
  • a particular embodiment of the invention is a pharmaceutical composition of the invention in which the NRG1 protein or peptide belongs to the following group: a) an amino acid sequence consisting of the human NRGl ⁇ amino acid sequence (SEQ ID
  • N02 an amino acid sequence consisting of the amino acid sequence NRGl ⁇ (SEQ ID N04), c) an amino acid sequence analogous to the sequence of a) and b), d) a fragment of any one of the sequences of a), b) and c), and e) an amino acid sequence comprising any sequence belonging to a) , b), c) and d).
  • Another particular object of the present invention is a pharmaceutical composition of the invention in which the compound or agent activating the development of neuron axons is a cell, preferably human, and more preferably a neuron, transformed by the sequence, construction or vector NRGl of the invention.
  • Another object of the invention is the use of the pharmaceutical composition of the invention, hereinafter use of the pharmaceutical composition of the invention, in a method of treatment or prophylaxis of a mammal, preferably a human being, affected by a disease, disorder or pathology that involves alterations of the development of axons of neurons consisting in the administration of said therapeutic composition in adequate dose that allows the recovery of the normal development of axons in neurons.
  • Another particular object of the present invention is the use of the pharmaceutical composition of the invention in a method of treating a disease or disorder that involves alterations in the development of axons of neurons that affect human beings, belonging, by way of illustration. and without limiting the scope of the invention, to the following group: traumatic spinal and brain injuries, neurodegenerative diseases, neuropsychiatric diseases and other neurological diseases related to GABAergic function.
  • a disease or disorder that involves alterations in the development of axons of neurons that affect human beings, belonging, by way of illustration. and without limiting the scope of the invention, to the following group: traumatic spinal and brain injuries, neurodegenerative diseases, neuropsychiatric diseases and other neurological diseases related to GABAergic function.
  • the present invention faces the problem of providing new therapeutic tools for the treatment of diseases that occur with alterations in the growth of neuron axons.
  • the present invention is based on the fact that the inventors have observed that isoforms of the Nrgl gene that contain the beta variant of the EGF domain (see SEQ ID N06 and N08) are capable of promoting the growth of axons and the development of branching neurons that express receptors for this factor, such as ErbB4 and ErbB3. This effect occurs both in long axon cells, such as thalamus-cortical neurons, and in short axon neurons, such as GABAergic interneurons of the cerebral cortex (Example 2).
  • NRGl EGF domain of NRGl (HERl ⁇ , in the case of the human form, see SEQ ID NO5 and NO6) is responsible for causing this effect, since a recombinant fragment of this domain is capable of producing the same effects that the complete NRGl sequence (Example 2), this fragment is found as part of the soluble NRGl (Ig-NRGl, see SEQ ID NO2 and NO4) or the membrane-anchored NRGl (CRD-NRGl, see SEQ ID NO14) .
  • Nrgl as a complete protein, and more specifically a peptide isolated from the EGF domain sequence of said protein, is an important factor for axonal development in different populations of nervous system neurons. It is therefore possible to think that this same factor could be used to regenerate nerve connections that depended on Nrgl / ErbB4 signaling by the therapeutic application of NRGl or its EGF domain (HERIb) either in its nucleotide or protein sequence form . This could be achieved through the development of strategies that allow expressing in the target neurons of the connections to regenerate a vector encoding Nrgl or its EGF domain (HERl ⁇ ) or using the protein or peptide as a therapeutic active compound to be applied directly.
  • NRGl or its EGF domain HERIb
  • an object of the present invention is the use of a compound that induces the activity of neuregulin 1 (hereinafter, NRGl), hereinafter used an activator compound of the present invention, for the preparation of a medicament or pharmaceutical composition for the treatment of a disease caused by an alteration of the axonal development of neurons of the nervous system.
  • NRGl neuregulin 1
  • the term "compound that induces NRG1 activity” refers to a molecule that mimics, increases the intensity or prolongs the duration of the developmental or regenerating activity of neuron axons of the NRG1 protein.
  • An activator compound may consist of a peptide, a protein or a nucleotide sequence, as well as those molecules that allow the expression of a nucleotide sequence encoding a protein with NRG1 activity.
  • disease caused by an alteration of the axonal development of neurons of the nervous system is refers to a disease, disorder or pathology belonging, among others by way of illustration and without limiting the scope of the invention, to the following group: chronic or acute traumatic spinal injuries, neurodegenerative diseases (such as Alzheimer's and Parkinson's disease) , neuropsychiatric diseases (such as schizophrenia and bipolar disorder), and other diseases of the nervous system such as epilepsy and autism.
  • neurodegenerative diseases such as Alzheimer's and Parkinson's disease
  • neuropsychiatric diseases such as schizophrenia and bipolar disorder
  • other diseases of the nervous system such as epilepsy and autism.
  • neurons refers to (i) neurons that are characterized by having long axons, for example, by way of illustration and without limiting the scope of the invention, thalamus-cortical neurons, which travel long distances to reach their target in the cerebral cortex, and (ii) to neurons that have short axons, for example, by way of illustration and without limiting the scope of the invention, inhibitory interneurons of the cerebral cortex, which have short axons that form local circuits in a restricted area of the cortex.
  • a particular object of the invention is the use of a compound that induces NRG1 activity based on the use of a nucleotide sequence, hereinafter NRG1 nucleotide sequence of the present invention, which allows the expression of a protein.
  • nucleotide sequence consisting of the nucleotide sequence encoding human NRGl ⁇ (SEQ ID NOl)
  • analogous is intended to include any nucleotide sequence that can be isolated or constructed based on the sequence shown herein, for example, by introducing conservative or non-conservative nucleotide substitutions. , including the insertion of one or more nucleotides, the addition of one or more nucleotides at any of the ends of the molecule or the deletion of one or more nucleotides at any end or within the sequence, and allowing the coding of a peptide or protein capable of mimicking the activity of the human protein sequence NRGl ⁇ (SEQ ID NO2), NRGlCC (SEQ ID NO4) or fragments thereof containing the EGF domain (HERl ⁇ and HERlCC, SEQ ID NO6 and SEQ ID NO8 , respectively).
  • NRG1-4 proteins belong to a family of factors present in several tissues that have different forms depending on the alternative splicing they suffer and that causes some forms to be anchored to the cell membrane (NRGl type III or CRD-NRGl forms) or that other soluble be secreted to the extracellular medium (NRGl type I / II or Nrgl-Ig forms) (Buonanno A, Fischbach GD (2001) Neuregulin and ErbB signaling receptor pathways in the nervous system. Curr Opin Neurobiol 11: 287-296). From the information described in the present invention and in the state of the art, a technician skilled in the art can isolate or construct a nucleotide sequence analogous to those described in the present invention for later use.
  • an analogous nucleotide sequence is substantially homologous to the nucleotide sequence discussed above.
  • the term "substantially homologous” means that the nucleotide sequences in question have a degree of identity of at least 30%, preferably of at least 85%, or more preferably of At least 95%.
  • Preferred forms of the nucleotide sequence to be used are the NRG1 nucleotide sequences or fragments thereof of human origin (see SEQ ID NO1, N03, N05 and N07)
  • nucleotide sequence refers to a sequence of DNA, cDNA or mRNA.
  • a particular embodiment of the present invention is the use of an activator compound of the invention where the nucleotide sequence NRG1 is constituted by SEQ ID NO1 (hNRGl ⁇ ), which codes for the human NRGl form of type I / II (Nrgl- Ig) which corresponds to Nrgl-typel ⁇ la (Acc. # NM_013956.1).
  • NRG1 is constituted by SEQ ID NO3 (hNRGl ⁇ ), which codes for the human NRGl form of type I / II (Nrgl- Ig) which corresponds to Nrgl-typel ⁇ la (Acc. # NM 013964.1).
  • NRG1 nucleotide sequence is constituted by a sequence that encodes the CRD-NRGl form of Ill ⁇ la type that corresponds to mouse Nrgl-typel ⁇ la ( Acc. # AY648975.1, see SEQ ID N013) or human (Acc # BC064587.1, see SEQ ID N015).
  • NRGl of c) analogous to the human form is the mouse sequence, and more specifically: the mNRGl ⁇ (SEQ ID N09), which codes for the NRGl form of mouse type I / II (Nrgl-Ig) corresponding to Nrgl -typel ⁇ la (Acc. #
  • nucleotide sequence of the NRG1 sequence of d) is a fragment comprising the coding sequence of the human EGF motif of neuregulin 1 (NRG1), preferably the nucleotide sequence of human herl ⁇ (SEQ ID NO5, Example 2) and the nucleotide sequence of human herl ⁇ (SEQ ID N07).
  • the NRG1 nucleotide sequence defined in section e) corresponds to an NRG1 gene construct that allows the expression of an NRGl protein.
  • This NRG1 gene construct of the invention can also comprise, if necessary and to allow a better isolation, detection or secretion outside the cell. of the expressed peptide, to a nucleotide sequence that encodes a peptide capable of being used for the purpose of isolation, detection or secretion of said peptide.
  • NRG1 genetic construct comprising, in addition to the NRG1 nucleotide sequence of the invention, any other nucleotide sequence encoding a peptide or peptide sequence that allows isolation, detection or the secretion outside the cell of the peptide expressed, for example, by way of illustration and without limiting the scope of the invention, a polyhistidine sequence (6xHis), a peptide sequence recognizable by a monoclonal antibody (for example, for its identification, or any other that serves to purify the resulting fusion protein by immunoaffinity chromatography: tag peptides such as c-myc, HA, E-tag) (Using antibodies: a laboratory manual. Ed. Harlow and David La ⁇ e
  • NRG1 nucleotide sequence and NRGl genetic construct described previously can be isolated and obtained by an expert by employing techniques widely known in the state of the art (Sambrook et al. "Molecular cloning, a Laboratory Manual 2 nd ed., CoId Sping Harbor Laboratory Press, NY, 1989 vol 1-3) Said nucleotide sequences can be integrated into a gene expression vector that allows regulation of the expression thereof under suitable conditions inside the cells.
  • NRG1 nucleotide sequence is an expression vector, hereinafter NRGl expression vector of the invention, which comprises an NRG1 nucleotide sequence or an NRGl genetic construct, described in the present invention, and which allows the expression of a protein or peptide capable of inducing axonal development in human neurons.
  • An example of a particular embodiment is the expression vectors elaborated in the present invention: pCMV2NRGl-SMDF (which allows the expression of the musdus CRD-Nrgl isoform), pCMV2NRGl-Igbeta (which allows the expression of the Ig isoform -Nrgl de Mus musculus) (see Examples, Material and Methods).
  • an expression vector comprises, in addition to the NGR1 nucleotide sequence or the NRG1 genetic construct described in the invention, a promoter that directs its transcription (eg, pT7, plac, ptrc, ptac, pBAD, ret, etc. .), to which it is operatively linked, and other necessary or appropriate sequences that control and regulate said transcription and, where appropriate, the translation of the product of interest, for example, transcription initiation and termination signals (tlt2, etc.) , polyadenylation signal, origin of replication, ribosome binding sequences (RBS), coding sequences of transcriptional regulators, (enhancers), transcriptional silencers (silencers), repressors, etc.
  • a promoter that directs its transcription eg, pT7, plac, ptrc, ptac, pBAD, ret, etc. .
  • other necessary or appropriate sequences that control and regulate said transcription and, where appropriate,
  • said vector is a plasmid or a viral vector.
  • the obtaining of said vector can be carried out by conventional methods known to those skilled in the art, as well as for the transformation of microorganisms and eukaryotic cells different widely known methods can be used - chemical transformation, electroporation, microinjection, etc. - described in various manuals [Sambrook, J., Fritsch, EF, and Maniatis, T. (1989). Molecular cloning: a laboratory manual, 2 nd ed. CoId Spring Harbor Laboratory, CoId Spring Harbor, NY].
  • One strategy could be to use lentivirus to infect the target cells, as it is already being tried in other types of therapies.
  • another particular object of the invention is the use of an inducing compound of the invention in which the activating molecule is a protein or peptide, hereinafter use of the NRG1 protein of the present invention, which exhibits axon-developing activity of human neurons, and comprising one or more amino acid sequences belonging to the following group: a) an amino acid sequence consisting of the human NRGl ⁇ amino acid sequence (SEQ ID NO: a) an amino acid sequence consisting of the human NRGl ⁇ amino acid sequence (SEQ ID
  • N02 an amino acid sequence consisting of the human NRGlOC amino acid sequence (SEQ ID N04), c) an amino acid sequence analogous to the sequence of a) and b), d) a fragment of any one of the sequences of a), b) and e), and e) an amino acid sequence comprising any sequence belonging to a), b), c) and d).
  • the term "analogous” is intended to include any amino acid sequence that can be isolated or constructed based on the sequence shown herein, for example, by introducing conservative or non-conservative amino acid substitutions. , including the insertion of one or more amino acids, the addition of one or more amino acids at any end of the molecule or the deletion of one or more amino acids at any end or inside the sequence, and that mimics the developmental activity of axons of neurons.
  • the NRGl protein belongs to the family of neuregulins (NRG1-4) as discussed above and is present in several organs. From the information described in the present invention and a technician skilled in the art field can isolate or construct an amino acid sequence analogous to those described in the present invention.
  • an analogous amino acid sequence is substantially homologous to the amino acid sequence discussed above.
  • the term "substantially homologous” means that the amino acid sequences in question have a degree of identity of at least 30%, preferably of at least 85%, or more preferably of At least 95%.
  • Another particular embodiment of the present invention is the use of an inducing compound of the invention in which the activating molecule is a protein NRGl whose amino acid sequence is constituted by human NRGl ⁇ type I / II (Nrgl-Ig) (SEQ ID N02).
  • NRGl whose amino acid sequence is constituted by the
  • Another particular embodiment of the present invention is the use of an inducing compound of the invention in which the activating molecule is an NRGl protein whose amino acid sequence is constituted by a CRD-NRGl form of Ill ⁇ la type corresponding to Nrgl-typel ⁇ la mouse (see SEQ ID NO14) or human (see SEQ ID NOl 6).
  • Another particular embodiment of the present invention is the use of an activating compound of the invention in which the activating molecule is an NRG1 protein whose NRGl amino acid sequence analogous to human form is mouse, and more specifically the mNRGlb (SEQ ID NOlO) and the mNRGla (SEQ ID NO12).
  • an inducing compound of the invention in which the activating molecule is a protein whose amino acid sequence of d) is a fragment comprising the coding sequence of the EGF motif of the neuregulins, preferably human HERl ⁇ (SEQ ID NO6) and human HERlCC (SEQ ID NO8) (Example 1).
  • NRG1 cells of the invention are used to express the nucleotide sequence, building or vector NRG1 expression of the invention - where the NRG peptide or protein of the invention can be adequately expressed - for the preparation of an inducing compound of the invention and its subsequent purification and application.
  • NRG1 cells of the invention genetically modified and comprising the nucleotide sequence, building or vector NRG1 expression of the invention - where the NRG peptide or protein of the invention can be adequately expressed - for the preparation of an inducing compound of the invention and its subsequent purification and application.
  • These cells can be transformed, infected or transcribed by said nucleotide sequences by genetic engineering techniques known to a person skilled in the art. [Sambrook, J., Fritsch, EF, and Maniatis, T. (1989).
  • a genetically transformed cell to express one of the NRG1 protein forms of the invention may allow the secretion of said NRG1 protein to the extracellular medium where it exerts its action, or the transformed cell itself may present said NRG1 protein in its membrane and participate as
  • Gene expression systems may or may not allow the integration of the new genetic material into the genome of the host cell.
  • both the nucleotide sequence, gene construct or NRG1 expression vector can be used as a medicament to protect human cells, preferably human neurons affected from an axonal alteration, in a method of treatment and prophylaxis of gene therapy of a being. human affected by a disease that involves neuronal disorders.
  • the NRG1 cells of the invention can be used as a medicament for the regeneration or implantation of tissues or cells in humans. Biopharmaceutical tools and gene therapy procedures are sufficiently known to a person skilled in the art in such a way that with the information described in the present invention they can be developed without undue effort.
  • a particular embodiment would be the use of a human cell transformed by these NRG1 nucleotide sequences, of different cell lines, preferably of the central nervous system and more preferably a neuron that can be used as regenerating cells of human tissues, and more preferably of the central nervous system .
  • proteins or peptides and the cells themselves can become biopharmaceuticals.
  • Another object of the present invention is a pharmaceutical composition or medicament for the treatment of diseases, disorders or pathologies that occur with alterations in the development of neuron axons, hereinafter pharmaceutical composition of the present invention, comprising a compound or inducing agent of the invention, in therapeutically effective amount together with, optionally, one or more pharmaceutically acceptable adjuvants and / or vehicles.
  • compositions are the adjuvants and vehicles known to those skilled in the art and commonly used in the elaboration of therapeutic compositions.
  • the term "therapeutically effective amount” refers to the amount of the agent or compound capable of developing axons of the neurons, calculated to produce the desired effect and, in general, will be determined, among other causes, by the characteristics of the compounds, including the age, condition of the patient, the severity of the alteration or disorder, and the route and frequency of administration.
  • said therapeutic composition is prepared in the form of a solid form or aqueous suspension, in a pharmaceutically acceptable diluent.
  • the therapeutic composition provided by this invention can be administered by any appropriate route of administration, for which said composition will be formulated in the pharmaceutical form appropriate to the route of administration chosen.
  • the administration of the therapeutic composition provided by this invention is carried out parenterally, orally, intraperitoneally, subcutaneously, etc.
  • Another particular object of the present invention is a pharmaceutical composition of the invention in which the compound or agent capable of developing axons of neurons belongs to the following group: sequence, genetic construction or NRGl vector that allow the expression of a protein or peptide NRGl of the invention.
  • a particular embodiment of the invention is a pharmaceutical composition of the invention in which the compound or agent capable of developing neuron axons is one or several NRG1 nucleotide sequences belonging to the following group: a) a nucleotide sequence consisting of the nucleotide sequence encoding the NRGl ⁇ (SEQ ID NOl), b) a nucleotide sequence consisting of the nucleotide sequence encoding the NRGlCC
  • NRG1 nucleotide sequence is constituted by SEQ ID NO1 (hNRGl ⁇ ), which codes for the human NRGl form of type I / II (Nrgl-Ig ) which corresponds to Nrgl-typel ⁇ la (Acc. # NM_013956.1).
  • NRG1 nucleotide sequence is constituted by SEQ ID N03 (hNRGlCC), which codes for the human NRGl form of type I / II (Nrgl-Ig ) which corresponds to Nrgl-typel ⁇ la (Acc. # NM_013964.1).
  • NRG1 nucleotide sequence is constituted by a sequence encoding the CRD-NRGl form of Ill ⁇ la type that corresponds to mouse Nrgl-typel ⁇ la (Acc. # AY648975.1, see SEQ ID N013) or human (Acc # BC064587.1, see SEQ ID N015).
  • nucleotide sequence of the NRG1 sequence of c) analogous to the human form is the mouse sequence, and more specifically: the mNRGl ⁇ (SEQ ID N09), which codes for the NRGl form of mouse type I / II (Nrgl-Ig) corresponding to Nrgl-typel ⁇ la (Acc. # AY648976.1), and - - the mNRGl ⁇ (SEQ ID NOIl), which codes for the form Mouse NRGl type I / II (Nrgl-Ig) corresponding to Nrgl-typel ⁇ la.
  • Another particular embodiment of the present invention is the pharmaceutical composition of the invention in which the NRG1 nucleotide sequence of d) is a fragment comprising the motif coding sequence.
  • Human EGF of Neuregulin 1 (NRGl), preferably the nucleotide sequence of human herl ⁇ (SEQ ID NO: 1
  • Example 2 Example 2 and the nucleotide sequence of human herl ⁇ (SEQ ID NO7).
  • Another particular embodiment of the present invention is a pharmaceutical composition of the invention in which the nucleotide sequence is an NRG1 expression vector.
  • Another particular object of the present invention is a pharmaceutical composition of the invention in which the compound or agent capable of recovering the development of neuron axons is a protein or a peptide encoded by the sequence, genetic construct or NRGl vector of the invention. .
  • a particular embodiment of the invention is constituted by a pharmaceutical composition of the invention in which the NRG1 protein or peptide belongs to the following group: a) an amino acid sequence consisting of the human NRGl ⁇ amino acid sequence (SEQ ID N02), b) an amino acid sequence consisting of the NRGl ⁇ amino acid sequence (SEQ ID N04) , c) an amino acid sequence analogous to the sequence of a) and b), d) a fragment of any one of the sequences of a), b) and e), and e) an amino acid sequence comprising any sequence belonging to a ), b), c) and d).
  • Another particular embodiment of the present invention is the pharmaceutical composition of the invention in which the amino acid sequence is an NRGl protein whose amino acid sequence is constituted by human NRGl ⁇ type I / II (Nrgl-Ig) (SEQ ID N02 ).
  • Another particular embodiment of the present invention is the pharmaceutical composition of the invention in which the amino acid sequence is an NRGl protein whose amino acid sequence is constituted by human NRGlCC type I / II (Nrgl-Ig) (SEQ ID NO4 ).
  • amino acid sequence is an NRGl protein whose amino acid sequence is constituted by a CRD-NRGl form of Ill ⁇ la type corresponding to mouse Nrgl-typel ⁇ la (see SEQ ID NO14) or human (see SEQ ID N016).
  • Another particular embodiment of the present invention is the pharmaceutical composition of the invention.
  • the amino acid sequence is an NRGl protein whose NRGl amino acid sequence of c) analogous to human form are mouse, and more specifically the mNRGl ⁇ (SEQ ID NO1) and the mNRGlCC (SEQ ID N012).
  • Another particular embodiment of the present invention is the pharmaceutical composition of the invention in which the amino acid sequence of d) is a fragment that comprises the coding sequence of the EGF motif of neuregulins, preferably human HERl ⁇ (SEQ ID NO6) and the human HERlCC (SEQ ID NO8) (Example 1).
  • Another particular object of the present invention is a pharmaceutical composition of the invention in which the compound or agent activating the development of neuron axons is a cell, preferably human, and more preferably a neuron, transformed by the sequence, construction or vector NRGl of the invention.
  • Another object of the invention is the use of the pharmaceutical composition of the invention, hereinafter use of the pharmaceutical composition of the invention, in a method of treatment or prophylaxis of a mammal, preferably a human being, affected by a disease, disorder or pathology that involves alterations of the development of axons of neurons consisting in the administration of said therapeutic composition in adequate dose that allows the recovery of the normal development of axons in neurons.
  • the pharmaceutical composition of the present invention can be used in a treatment method in isolation or in conjunction with other pharmaceutical compounds.
  • Another particular object of the present invention is the use of the pharmaceutical composition of the invention in a method of treating a disease or disorder that involves alterations of the development of axons of neurons that affect human beings, belonging, by way of illustration and without limiting the scope of the invention, to the following group: traumatic spinal and brain injuries, neurodegenerative diseases, neuropsychiatric diseases and other neurological diseases related to function GABAergic.
  • Another particular embodiment of the present invention is the use of the pharmaceutical composition of the invention in a method of treating a neurodegenerative disease belonging to the following group: multiple sclerosis, Alzheimer's disease and Parkinson's.
  • Another particular embodiment of the present invention is the use of the pharmaceutical composition of the invention in a method of treating a neuropsychiatric disease belonging to the following group: schizophrenia and bipolar disorder.
  • Another particular embodiment of the present invention is the use of the pharmaceutical composition of the invention in a method of treatment of other neurological diseases belonging to the following group: epilepsy and autism.
  • Ig-NRGl promotes the growth of the thalamus-cortical axons (TCAs).
  • TCAs thalamus-cortical axons
  • A Expression of Ig-Nrgl in the cerebral cortex (NCx) to E13.5.
  • B, C Immunohistochemistry for ⁇ lII-Tubulin, a neuronal marker, showing explants of the dorsal thalamus (dTh) of E13.5 embryos after 96 hours in culture, facing aggregates of control COS cells (B) or transcribed with Ig-Nrgl (C) .
  • the boxes show the expression of GFP in the transfected COS cells.
  • MGE lateral ganglionic eminence
  • LGE lateral ganglionic eminence
  • Axons are labeled with an antibody against ⁇ lll-Tubulin, a neuronal protein.
  • FIG. 3 Expression of Nrgl and its ErbB4 receptor in the postnatal cerebral cortex.
  • A, A ' Cross sections through the telencephalon of a 30 day postnatal mouse (P30) showing the expression of the CRD-Nrg-1 isoform by in situ hybridization.
  • A Panoramic view of the strong expression of CRD-Nrg-1 in the neocortex (NCx) and hippocampus (HC).
  • a ' Detail of the cortical region bounded by a dashed line in A.
  • NRGl stimulates the growth of axons and their collaterals in cortical interneurons.
  • A Experimental paradigm used to demonstrate the role of NRGl in the axonal development of GABAergic interneurons.
  • B Image of an interneuron in culture after being treated with control supernatant.
  • Example 1 The role of NRGl in the development of axons of neurons of the nervous system.
  • thalamus-cortical neurons which are characterized by having long axons that travel long distances to reach their target in the cerebral cortex
  • cortex inhibitory interneurons brain which have short axons that form local circuits in a restricted area of the cortex.
  • Example 2.- NRGl is a potent stimulator of the growth of short axons.
  • Nrgl is a potent stimulator of the growth of long axons, as is the case of TCAs, it was proposed to investigate the role that this molecule could have in the development of short axons, as is the case of interneuron axons cortical
  • the distribution of Nrgl in the postnatal cortex seems to indicate that it is found mainly in projection neurons, also called pyramidal cells.
  • NRG1 isoforms used, as well as the NRGl EGF domain (HERl ⁇ ), were able to induce phosphorylation of ErbB3 and ErbB4 receptors in cell line assays (data not shown), which shows that the The activity of these proteins and peptides is mediated, at least in part, through these receptors.
  • C0S7 cells were transiected with a control plasmid or with a Nrgl-encoding plasmid (mouse CRD-Nrgl form (SEQ ID NO13)), or with the EGF motif of human Nrgl ⁇ (SEQ ID NO5). and its supernatant was collected after 48 hours in culture.
  • mice Animals The experiments have been carried out using wild mice from two different genetic backgrounds (CD1 and C57b / 6), and transgenic mice with generalized GFP expression or with GFP expression under the GAD65 promoter (GAD65: GFP). All the lines were maintained in the Animal Experimentation Service of the Miguel Hernández University of Elche in accordance with the regulations in force in Spain and the guidelines of the European Union.
  • Organotypic sliced cultures were performed as described in detail previously (Anderson SA, Eisenstat DD, Shi L, Rubenstein JLR (1997) Interneuron migration from basal forebrain to neocortex: dependence on DIx genes. Science 278: 474- 476).
  • the slices were grown on polycarbonate membranes for culture (8 ⁇ m pore size; Corning Costar) in culture plates with 1 ml of medium (Neurobasal / B-27 [Life Technologies] supplemented with glutamine, 5% horse serum and pen / strep). In these trials, the TCAs begin to grow after 36 hours of growth; The slices were grown for 72-96 hours.
  • the aggregates of COS7 cells are prepared by diluting the transcribed cells with Matrigel as described previously (Flames N, Long JE, Garratt AN, Fischer TM, Gassmann M, Birchmeier C, Lai C, Rubenstein JL, Marin 0 (2004) Short- and long-range attraction of cortical GABAergic interneurons by neuregulin-1. Neuron 44: 251-261).

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Abstract

La présente invention décrit comment des composés thérapeutiques présentant différents isoformes du gène Nrg1 contenant le variant bêta du domaine EGF sont capables de promouvoir la croissance des axones et le développement de ramifications de neurones exprimant des récepteurs pour ce facteur, tels que ErbB4 et ErbB3. L'invention concerne également l'utilisation desdits composés dans l'élaboration de compositions pharmaceutiques et leurs application dans le traitement de maladies causées par des altérations du développement axonal, telles que des lésions de la moelle épinière ou encore des maladies neurodégénératives et psychiatriques.
PCT/ES2007/070068 2006-04-05 2007-04-02 Utilisation de composés induisant le développement axonal des neurones, compositions thérapeutiques contenant ces composés et leurs applications WO2007113366A1 (fr)

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Cited By (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN105561299A (zh) * 2007-11-16 2016-05-11 意识-Nrg公司 经翻译后修饰的有活性的可溶性神经调节蛋白同种型
CN108277272A (zh) * 2018-04-09 2018-07-13 重庆医科大学附属第医院 一种辅助癫痫诊断的标志物及其检测试剂盒
US11426447B2 (en) * 2016-04-19 2022-08-30 Leibniz-Institut Fur Alternsforschung—Fritz-Lipmann-Institut E.V. (Fli) Neuregulin for the treatment of tumors of the nervous system

Families Citing this family (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2008148023A2 (fr) * 2007-05-23 2008-12-04 Medical College Of Georgia Research Institute, Inc. Compositions et procédés pour traiter des troubles neurologiques
KR101186218B1 (ko) * 2009-08-21 2012-10-08 경희대학교 산학협력단 히레귤린 베타1의 ｅｇｆ-성 도메인 펩타이드의 용도

Citations (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2001026607A2 (fr) * 1999-10-08 2001-04-19 Uab Research Foundation Isoformes de variant d'epissage de neureguline smdf et ggf et utilisations de celles-ci
US20030129688A1 (en) * 1997-02-10 2003-07-10 Genentech, Inc. Heregulin variants
US20050208527A1 (en) * 2000-02-28 2005-09-22 Decode Genetics Ehf. Human schizophrenia gene

Patent Citations (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US20030129688A1 (en) * 1997-02-10 2003-07-10 Genentech, Inc. Heregulin variants
WO2001026607A2 (fr) * 1999-10-08 2001-04-19 Uab Research Foundation Isoformes de variant d'epissage de neureguline smdf et ggf et utilisations de celles-ci
US20050208527A1 (en) * 2000-02-28 2005-09-22 Decode Genetics Ehf. Human schizophrenia gene

Non-Patent Citations (3)

* Cited by examiner, † Cited by third party
Title
ESPER R.M. ET AL.: "Neuregulins: versatile growth and differentiation factors in nervous system development and human disease", BRAIN RES. REV., vol. 51, no. 2, August 2006 (2006-08-01), pages 161 - 175, XP005508484 *
FLAMES N. ET AL.: "Short- and long-range attraction of cortical GABAergic interneurons by neuregulin-1", NEURON., vol. 44, no. 2, 2004, pages 251 - 261 *
HO W.-H. ET AL.: "Sensory and motor neuron-derived factor. A novel heregulin variant highly expressed in sensory and motor neurons", J. BIOL. CHEM., vol. 270, no. 24, 1995, pages 14523 - 14532, XP002003223 *

Cited By (5)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN105561299A (zh) * 2007-11-16 2016-05-11 意识-Nrg公司 经翻译后修饰的有活性的可溶性神经调节蛋白同种型
US10350269B2 (en) 2007-11-16 2019-07-16 Mind-Nrg Sarl Method of effecting neuroprotection using soluble Neuregulin isoforms
US10813978B2 (en) 2007-11-16 2020-10-27 Mind-NRG Sàrl Method of effecting neuroprotection using soluble neuregulin isoforms
US11426447B2 (en) * 2016-04-19 2022-08-30 Leibniz-Institut Fur Alternsforschung—Fritz-Lipmann-Institut E.V. (Fli) Neuregulin for the treatment of tumors of the nervous system
CN108277272A (zh) * 2018-04-09 2018-07-13 重庆医科大学附属第医院 一种辅助癫痫诊断的标志物及其检测试剂盒

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