WO2005082135A1 - Promoteur de regeneration d'axone - Google Patents

Promoteur de regeneration d'axone Download PDF

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Publication number
WO2005082135A1
WO2005082135A1 PCT/JP2005/002924 JP2005002924W WO2005082135A1 WO 2005082135 A1 WO2005082135 A1 WO 2005082135A1 JP 2005002924 W JP2005002924 W JP 2005002924W WO 2005082135 A1 WO2005082135 A1 WO 2005082135A1
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pkc
axon
regeneration
inhibitor
myelin
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PCT/JP2005/002924
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English (en)
Japanese (ja)
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Toshihide Yamashita
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Bioclues, Inc.
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Priority to JP2006510433A priority Critical patent/JPWO2005082135A1/ja
Publication of WO2005082135A1 publication Critical patent/WO2005082135A1/fr

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    • 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
    • C07K14/705—Receptors; Cell surface antigens; Cell surface determinants
    • 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
    • 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
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P43/00—Drugs for specific purposes, not provided for in groups A61P1/00-A61P41/00
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N2333/00—Assays involving biological materials from specific organisms or of a specific nature
    • G01N2333/90—Enzymes; Proenzymes
    • G01N2333/91—Transferases (2.)
    • G01N2333/912—Transferases (2.) transferring phosphorus containing groups, e.g. kinases (2.7)
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N2500/00—Screening for compounds of potential therapeutic value
    • G01N2500/04—Screening involving studying the effect of compounds C directly on molecule A (e.g. C are potential ligands for a receptor A, or potential substrates for an enzyme A)

Definitions

  • the present invention relates to an axon regeneration-promoting agent that can promote the regeneration of axons of nerve cells, particularly nerve cells of the central nervous system.
  • Non-Patent Document 1 myelin-binding glycoprotein (MAG) and oligodendrocyte-myelin glycoprotein ( ⁇ Mgp) have been identified as three major inhibitors of central nervous system regeneration. Nogo was identified as a corresponding antigen of the monoclonal antibody IN-1 (Non-patent Documents 2-4).
  • Non-Patent Document 58 which is known to play an important role in the formation and maintenance of the myelin sheath, was found to inhibit the growth of certain neuronal axons ( Non-patent documents 9, 10).
  • OMgp Non-Patent Document 11
  • Nogo, MAG and Mgp are known to bind to NgR using p75 as a co-receptor, suggesting that they share a common signaling pathway (Non-Patent Document 14-1 17).
  • Non-Patent Document 1 S. David, A.J.Aguayo, Science 214, 931-3 (Nov 20, 1981).
  • Non-Patent Document 2 M. S. Chen et al "Nature 403, 434-9 (Jan 27, 2000).
  • Non-Patent Document 3 T. GrandPre, F. Nakamura, T. Vartanian, S.M.Strittmatter, Nature 403, 439-44 (Jan 27, 2000).
  • Non-Patent Document 4 p. Caroni, ME Schwab, Neuron 1, 85-96 (Mar, 1988).
  • Non-Patent Document 7 N. Fujita et al., J Neurosci 18, 1970-8 (Mar 15, 1998).
  • Non-Patent Document 8 J. Marcus, J. L. Dupree, B. Popko, J Cell Biol 156, 567-77 (Feb 4, 2002).
  • Non-Patent Document 9 G. Mukhopadhyay, P. Doherty, F.S.Walsh, P.R.Crocker, MT Filbin, Neuron 13, 757-67 (Sep, 1994).
  • Non-Patent Document 10 and McKerracher et al., Neuron 13, 805-11 (Oct, 1994).
  • Non-Patent Document 11 D.D.Mikol, K. Stefansson, J Cell Biol 106, 1273-9 (Apr, 1988).
  • Non-Patent Document 12 V. Kottis et al., J Neurochem 82, 1566-9 (Sep, 2002).
  • Non-Patent Document 13 K. C. Wang et al., Nature 417, 941-4 (Jun 27, 2002).
  • Non-Patent Document 14 ⁇ C. Wang, J.A. Kim, R. Sivasankaran, R. Segal, Z. He, Nature 420, 74-8 (Nov 7, 2002).
  • Non-Patent Document 15 M. Domeniconi et al, Neuron 35, 283-90 (Jul 18, 2002).
  • Non-Patent Document 16 A. E. Fournier, T. GrandPre, S.M.Strittmatter, Nature 409, 341-6 (Jan 18, 2001).
  • Non-Patent Document 17 T. Yamashita, ⁇ . Higuchi, ⁇ . Tohyama, J Cell Biol 157, 565-70 (May 13, 2002).
  • Non-Patent Document 18 U. Bartsch et al "Neuron 15, 1375-81 (Dec, 1995).
  • Non-Patent Document 20 J. E. Kim, S. Li, T. GrandPre, D. Qiu, S. M.Strittmatter, Neuron 38, 187-99 (Apr 24, 2003).
  • Non-Patent Document 21 B. Zheng et al., Neuron 38, 213-24 (Apr 24, 2003).
  • Non-Patent Document 22 M. Simonen et al., Neuron 38, 201-11 (Apr 24, 2003).
  • Non-Patent Document 23 X. Song et al., J Neurosci 24, 542-6 (Jan 14, 2004).
  • Non-Patent Document 24 Lin, W.W., Wang, C.W. & Chuang, D.M.J.Neurochem. 68, 2577-2586 (1997).
  • Non-Patent Document 25 Eichholtz, T., et al. 1993. J. Biol. Chem. 28, 1982; Ward, N.E., and OAiBrian, C.A. 1993. Biochemistry 32, 11903.
  • Non-patent document 27 Seewald, S. et al. (1999) Am. J. Hypertens. 12: 532-537
  • Non-patent document 28 Sveltov, S., and Nigami, S. 1993. Biochim. Biophys. Acta 1177, 75
  • Non-Patent Document 29 Ku, W.-C, et al. 1997. Biochem. Biophys. Res. Commun. 241, 730
  • Non-Patent Document 30 Doherty, P. et al., 1990. Nature 343: 464-466 Disclosure of the Invention
  • An object of the present invention is to provide a novel axon regeneration-promoting agent that can promote regeneration of central axons.
  • MAG and Nogo which had been considered to be substances that inhibit axonal regeneration, were reversed in the state where the function of conventional protein kinase C was inhibited.
  • a function to promote axonal regeneration The present inventors have found that a novel protein kinase c inhibitor is effective in promoting axonal regeneration, and completed the present invention.
  • the present invention provides an axon regeneration promoter containing a conventional protein kinase C inhibitor as an active ingredient.
  • the axons are those of the central nervous system.
  • said conventional protein kinase C is proteinase C-hi, ⁇ 1, 2, ⁇ or ⁇ .
  • said conventional oral tin kinase C inhibitor is a cell permeable PKC inhibitor 20-28 or Go6976.
  • the axon regeneration-promoting agent of the present invention is a myelin-binding glycoprotein, Nogo peptide or myelin, or a mutant thereof, wherein the conventional protein kinase C is inhibited by the inhibitor. And a fusion protein containing these or a mutant thereof and exhibiting an axonal regeneration-promoting effect. More preferably, the axon regeneration promoter of the present invention contains myelin-binding glycoprotein, Nogo peptide or myelin, or a fusion protein containing these.
  • the present invention relates to a method for identifying a candidate substance for promoting axonal regeneration, comprising contacting a test substance with a conventional PKC, wherein the test substance inhibits the function of the conventional PKC.
  • Providing a method comprising determining whether
  • axon regeneration-promoting agent capable of promoting regeneration of axons of the central nervous system.
  • the axon regeneration-promoting agent of the present invention is effective for regeneration of axons of the central nervous system, and is expected to greatly contribute to treatment of patients suffering from damage to the central nervous system such as the spinal cord.
  • Fig. 1 shows the relative activity of PKC when cultured cerebellar granule neurons were treated with MAG for 10, 30 or 1 hour. PKC activity is indicated by the amount of phosphorylated PKC- ⁇ relative to the amount of PKC-hi. Relative values are expressed as a ratio to PKC activity at time zero. The results show the average soil standard deviation from three experiments. An asterisk indicates that it is statistically significant (p 0.01 in Student's t-test).
  • Figure 2 shows the axon length of cerebellar granule neurons in the presence of the components shown. You. In the presence of PKCI, MAG-Fc and Nogo peptide each stimulated axonal growth. Data show mean soil standard deviation. An asterisk indicates that it is statistically significant (P 0.01 in Student's t-test).
  • Figure 3 shows the results of growth cone collapse atsushi. Data indicate mean ⁇ standard deviation. An asterisk indicates that it is statistically significant (p 0.01 in Student's t-test).
  • FIG. 4 shows the results of affinity precipitation of RhoA in cerebellar granule neurons.
  • MAG-Fc and Nogo peptide activate RhoA in the presence or absence of PKCI Best mode for carrying out the invention
  • axon regeneration promoter of the present invention are those which contain conventional protein kinase C inhibitor as an active ingredient (hereinafter, the "protein kinase ⁇ ""1 «:?”Abbreviated child and force s is there).
  • protein kinase ⁇ protein kinase C inhibitor
  • PKC-a, PKC-J31, PKC-2, PKC- ⁇ both nonpatent literature 24
  • Inhibitors of PKC-II and PKC- ⁇ atypical PKCs other than the conventional PKC, have no axonal regeneration promoting effect.
  • PKCI PKC inhibitors
  • the cell permeable PKC inhibitor 20-28, Go 6976 and the cell permeable PKC inhibitor 19-27 are preferred, and the cell permeable PKC inhibitor 20-28 and Go 6976 are particularly preferred.
  • These PKCIs can be used alone or in combination of two or more.
  • the cell permeable PK and inhibitor 20-28 N-Myr- Phe- Ala- Arg- Arg- Lys- Gly- Ala- Leu- Arg-Gln- NH (SEQ ID NO: 5)
  • the axon regeneration-promoting agent of the present invention may contain a monoclonal antibody capable of binding to conventional PKC and inhibiting its function as a conventional protein kinase C inhibitor.
  • a monoclonal antibody capable of binding to conventional PKC and inhibiting its function as a conventional protein kinase C inhibitor.
  • the animal is immunized using conventional PKC as a sensitizing antigen and the resulting immune cells are removed and fused with myeloma cells according to methods well known in the art. Then, clone the hybridoma producing the antibody.
  • Monoclonal antibodies can be prepared from culture supernatants obtained by culturing hybridomas or from mouse ascites fluid after intraperitoneal administration of the hybridomas to mice.
  • recombinant antibodies in addition to the antibodies produced by humans and hybridomas, recombinant antibodies, chimeric antibodies, CDR-grafted antibodies, fragments of these antibodies, and the like may also be used as the complementary PKC inhibitors in the present invention. Can be.
  • the recombinant antibody is obtained by cloning cDNA encoding the antibody from a hybridoma that produces a monoclonal antibody that binds to conventional PKC, and introducing it into an expression vector to transform animal cells, plant cells, and the like. It is possible to produce by transforming and cultivating this transformant.
  • a chimeric antibody is defined as a heavy chain variable region and a light chain variable region of an antibody derived from an animal (eg, a mouse), and a heavy chain constant region and a light chain constant region of an antibody derived from another animal (eg, a human). Area force is composed of antibodies.
  • a chimeric antibody clones the cDNAs encoding the heavy and light chain variable regions of the antibody from a hybridoma producing a monoclonal antibody that binds to conventional PKC, while retaining the weight of antibodies derived from other animals. It can be produced recombinantly by cloning the cDNAs encoding the chain constant region and the light chain constant region, combining them, inserting them into an appropriate expression vector, and expressing them in host cells.
  • a CDR-grafted antibody is an antibody obtained by grafting the complementarity-determining region (CDR) of an antibody of one animal (eg, a mouse) to the complementarity-determining region of an antibody of another animal (eg, a human).
  • the gene encoding the CDR-grafted antibody is based on the gene sequences of the heavy chain variable region and light chain variable region of an antibody cloned from a hybridoma that produces a monoclonal antibody that binds to conventional PKC. It can be obtained by designing the sequence and replacing it with the corresponding CDR sequence in a vector containing the genes encoding the heavy and light chain variable regions of antibodies from other animals.
  • Examples of antibody fragments that can bind to conventional PKC include Fab, F (ab ') 2, Fab', scFv, and diabody. These antibody fragments can be used to transform the above-mentioned anti-conventional PKC monoclonal antibody of the present invention into host cells by the ability to treat it with an enzyme such as papain or trypsin, or by introducing an expression vector incorporating a gene encoding these into a host cell. By obtaining, the manufacturing power can be achieved.
  • the axon regeneration-promoting agent of the present invention includes antisense oligonucleotides, ribozymes, and molecules that cause RNA interference (RNAi) (eg, dsRNA, siRNA, shRNA, and miRNA) as conventional protein kinase C inhibitors. ) May be included.
  • RNAi RNA interference
  • These nucleic acids can bind to and inhibit the expression of a conventional PKC gene or mRNA encoding a conventional PKC.
  • General methods for controlling gene expression using antisense, ribozyme and RNAi techniques, or gene therapy methods for expressing exogenous genes in this manner, are well known in the art.
  • the antisense oligonucleotide refers to a nucleic acid molecule having a sequence complementary to mRNA encoding conventional PKC or a derivative thereof. Antisense oligonucleotides specifically hybridize to mRNA and inhibit protein expression by inhibiting transcription and Z or translation. Binding may be by Watson'Crick or Houdasteen base pair complementarity or by triplex formation.
  • a ribozyme refers to the RNA structure of one or more RNAs having catalytic properties. Ribozymes generally exhibit endonuclease, ligase or polymerase activity. Various secondary ribozymes are known, such as hammerhead and hairpin ribozymes.
  • RNA interference (RNAi) refers to a method of silencing a target gene using double-stranded RNA molecules.
  • parenteral administration is preferred, and it is particularly preferable to inject directly into the nerve injury site.
  • the dose is appropriately selected according to the type of PKCI, the administration route, the degree of nerve damage, and the like.
  • the dose of PKCI per adult site per injury site per day for Go 6976 is usually O.Olmg lmg, preferably 0.05mg-0.5mg.
  • the dose is usually about 10 times the above dose.
  • PKCI can be administered as it is, but is usually formulated using a carrier used in medicine.
  • a carrier used in medicine any of those commonly used in the field of preparation can be used.
  • physiological saline or phosphate buffered saline is preferably used for preparing an injection.
  • commonly used additives such as emulsifiers and osmotic pressure regulators may be included.
  • the axon regeneration-promoting agent of the present invention is effective for promoting the regeneration of axons of nerve cells, particularly those of the central nervous system such as the spinal cord. Therefore, it can be used to treat central nervous system injuries caused by accidents.
  • mouse neurons were extracted and tested. Therefore, experiments were performed with the addition of MAG, Nogo, or myelin. For nerve fibers in vivo, these are contained in the nerve sheath. Therefore, it is not necessary to add MAG, Nogo or myelin separately to the axon regeneration promoter.
  • MAG, Nogo or myelin may be separately added to the axon regeneration promoter.
  • MAG, Nogo and myelin themselves are well known and readily available.
  • the nucleotide sequence of the human MAG gene and the amino acid sequence encoded thereby are shown in SEQ ID NOs: 1 and 2 (GenBank Accession No. AC002132).
  • the nucleotide sequence of the human Nogo gene and the amino acid sequence encoded thereby are shown in SEQ ID NOs: 3 and 4, respectively ( GenBank Accession No. AY102279).
  • the gene and amino acid sequences of MAG and Nogo of non-human mammals are also known.
  • a polypeptide having a physiological activity may be a mutant in which a small number of amino acids constituting the amino acid sequence have been substituted or deleted, or a small number of amino acids have been inserted. It is widely known that the physiological activity may be maintained.
  • MAG, Nogo or myelin is added to the axon regeneration-promoting agent of the present invention, these variants are those that exert an axonal regeneration-promoting effect when PKC is inhibited by the above inhibitor. May be added.
  • Such variants are preferably those having an amino acid sequence having at least 80%, preferably at least 90%, more preferably at least 95% identity to the amino acid sequence of the native polypeptide, particularly substitutions.
  • one or several amino acids are deleted or inserted.
  • amino acid sequence identity can be easily determined using BLAST, a software widely used in this field.
  • BLAST is available to anyone on the NCBI (National Center for Biotechnology Information) homepage, and identity can be easily checked using default parameters.
  • the 20 types of amino acids that make up the natural protein are the neutral amino acids He, Val, Leu, Ala, Met, and Pro with low-polarity side chains, and the neutral amino acids with hydrophilic side chains (Asn and Gin). , Thr, Ser, Tyr, Cys), acidic amino acids (Asp, Glu), basic amino acids (Arg, Lys, His), aromatic amino acids (Phe, Tyr, TYp) Can be divided.
  • fusion proteins containing MAG, Nogo, or myelin or their mutants usually maintain the biological activity of these polypeptides. You can also.
  • a fusion protein of MAG and the Fc region of an immunoglobulin is used, but such a fusion protein may be added to the axon regeneration promoter of the present invention.
  • MAG MAG, Nogo or myelin
  • a natural substance derived from the animal species to be administered meaning that it contains no mutation (however, it may be produced by genetic engineering techniques)
  • Particularly preferred is a naturally-occurring protein which is preferred by a fusion protein containing it.
  • the concentration of MAG which is an axon regeneration promoter, is usually 01000 ⁇ , preferably 050 ⁇ , and the concentration of Nogo is usually 0-10 ⁇ , preferably 0-0.5.
  • the concentration of myelin is usually 0 lOOng / ⁇ , preferably 0-lOng / ⁇ .
  • the present invention provides a method for identifying a candidate substance for promoting axonal regeneration.
  • the method involves contacting the test substance with a conventional PKC and determining whether the test substance inhibits the function of the conventional PKC.
  • the functions of conventional PKC include both the enzymatic activity of conventional PKC to phosphorylate its substrate protein and the ability of conventional PKC to bind ligand.
  • the enzymatic activity of conventional PKC can be easily measured using any of the various kinase activity assays known in the art. By comparing the kinase activity in the presence and absence of the test substance, it is possible to determine force to inhibit the enzymatic activity of the test substance force S conventional p KC. Also
  • the ability of a conventional PKC to bind its corresponding ligand can be determined by binding assays well known to those skilled in the art, such as, but not limited to, gel shift assay, radiolabel competition assay, chromatography fractionation, and the like. it can.
  • Substances identified as inhibiting conventional PKC function by these assays are considered to be candidate axon regeneration promoters.
  • the candidate substance is then axon-measured by measuring and comparing neuronal axon growth in the presence and absence of the candidate substance using Atsey methods known in the art. It can be determined whether or not it has a regeneration promoting effect.
  • the PKC assay was performed using a PepTag assay kit (trade name, Promega, Madison, Wis., USA), which is a PKC-based detection kit without radioactivity.
  • Serum-starved cerebellar granule cells were cultured with MAG_Fc (a fusion protein of MAG and immunoglobulin Fc region, available from Sigma, St. Louis, Mo., USA) (25 / ig / ml) and Nogo peptide X Alpha Diagnostic, San Antonio, Texas, USA, 4 ⁇ ⁇ ) was applied. Each sample was incubated for 30 minutes at 30 ° C with PKC substrate ⁇ -marked Tag C1 peptide (2 / ig).
  • the samples were separated on a 0.8% agarose gel at 100 V for 15 minutes.
  • the phosphorylated peptide substrate moved to the anode (+), and the non-phosphorylated peptide substrate moved to the cathode (-).
  • the gel was photographed on a transilluminator.
  • a phosphate-specific antibody was used for the measurement of phosphorylated PKC. 10 mM Tris-HCl [pH 7.5], 150 mM NaCl, 10% glycerol, 100 ⁇ M sodium orthovanadate, 50 mM sodium fluoride, 5 mM sodium pyrophosphate, 1 mM EGTA and protease inhibitor mixture ( The cells were lysed with 0.1% NP_40 (trade name, nonionic surfactant lj) containing Roche Biochemicals). The cell lysate was subjected to SDS-PAGE, and Western blot analysis was performed by a conventional method.
  • NP_40 trade name, nonionic surfactant lj
  • Polyclonal anti-phospho PKC-H (Ser657 phosphorylated, Upstate Biotech, Charlottesville, VA, USA), polyclonal anti-phospho PKC-/ ⁇ (Cell Signaling Tech) and polyclonal anti-PKC-a antibody (Santa Cruz Biotech) Manufactured).
  • cerebellar neurons cerebellum from two 7-day-old rats was They were combined in a 0.025% trypsin solution, crushed, and incubated at 37 ° C for 10 minutes. DMEM medium containing 10% FCS was added, and the cells were collected by centrifugation at 800 ⁇ m. Neurons were plated in Sato medium (Non-Patent Document 30) on chamber slides coated with poly-L_lysine. For the growth assay, plated cells were incubated for 24 hours, fixed with 4% (w / v) paraformaldehyde, and monoclonal antibodies (Tujl, 4) were used to recognize ⁇ -tubulin III protein specific to neurons.
  • Tris containing 1% Triton X_100 (trade name), 0.5% sodium deoxycholate, 0.1% SDS, 500 mM NaCl and 10 mM MgCl, 10 ⁇ g / ml of leptin and 10 ⁇ g / ml of aprotune (pH 7.
  • the cells were lysed in 5).
  • the cell lysate is clarified by centrifugation at 13000g at 4 ° C for 10 minutes, and the supernatant is combined with 20 ⁇ g of GST-Rho binding region of Rhotekin beads (trade name, manufactured by Upstate Biotech) at 4 ° C for 45 minutes. Incubated.
  • the beads were washed four times with a washing buffer (1% Triton X_100 (trade name), 150 mM NaCl, 10 mM MgCl, 50 mM Tris (pH 7.5) containing 10 ⁇ g / ml each of leptin and aprotune).
  • a washing buffer 1% Triton X_100 (trade name), 150 mM NaCl, 10 mM MgCl, 50 mM Tris (pH 7.5) containing 10 ⁇ g / ml each of leptin and aprotune.
  • Monoclonal antibody against RhoA (Santa Cruz Biotech, Santa Cruz, CA, USA) Bound Rho was detected by the Western blot used.
  • MAG-Fc and Nogo peptide are not available in the presence of the cell permeable PKC inhibitor 20-28 (indicated as “PKCI” in Figure 2), which is a PKC- ⁇ and -inhibitor. , Which dramatically stimulated axonal growth (Figure 2), but PKCI itself had no axonal growth effect (Figure 2) Axonal growth induced by MAG-Fc or Nogo Similar results were obtained with Go6976, an inhibitor of PKC-a and ⁇ (Fig. 2), but not with the Rho kinase inhibitor. is there
  • Rho has been shown to be a key signal molecule in axonal growth inhibition (Non-Patent Documents 17 and 14). Therefore, RhoA activity in neurons was measured. RhoA was activated 30 minutes after the addition of MAG-Fc or Nogo to cerebellar neurons of 7-day-old rats (Fig. 4). PKCI had no effect on Rho activity induced by MAG-Fc or Nogo peptide. Thus, promotion of axonal growth of cerebellar neurons by inhibiting PKC is not mediated by blocking RhoA activity, indicating that conventional PKC is not upstream of RhoA. I have.
  • the axon regeneration-promoting agent of the present invention is effective for regeneration of a damaged central nervous system, and is useful as a therapeutic agent for patients having a damaged central nervous system.

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Abstract

Promoteur de régénération d'axone qui contient un inhibiteur de kinase C de protéine traditionnelle en tant qu'ingrédient actif. Ce promoteur de régénération d'axone dans le système nerveux central contribue par exemple au traitement pour un patient souffrant d'un traumatisme du système nerveux central à la suite d'un accident de la route, d'un désordre cérébrovasculaire etc. etc.
PCT/JP2005/002924 2004-02-26 2005-02-23 Promoteur de regeneration d'axone WO2005082135A1 (fr)

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Citations (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US20030176423A1 (en) * 2002-03-14 2003-09-18 Children's Medical Center Corporation Axon regeneration with pkc inhibitiors
WO2004087744A2 (fr) * 2003-03-28 2004-10-14 Intellectual Property Consulting Inc. Composition et methode de regeneration nerveuse

Patent Citations (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US20030176423A1 (en) * 2002-03-14 2003-09-18 Children's Medical Center Corporation Axon regeneration with pkc inhibitiors
WO2004087744A2 (fr) * 2003-03-28 2004-10-14 Intellectual Property Consulting Inc. Composition et methode de regeneration nerveuse

Non-Patent Citations (3)

* Cited by examiner, † Cited by third party
Title
EICHHOLTZ T. ET AL: "A Myristoylated Pseudosubstrate Peptiode, a Novel Protein Kinase C Inhibitor", THE JOURNAL OF BIOLOGICAL CHEMISTRY, vol. 268, no. 3, 1993, pages 1982 - 1986, XP002988788 *
PRANG P. ET AL: "Regeneration of enthorthinal fibers in mouse slice cultures is age dependant and can be stimulated by NT-4, GDNF, and modulators of G-proteins and protein kinase C", EXPERIMENTAL NEUROLOGY, vol. 169, no. 1, 2001, pages 135 - 147, XP002973622 *
TOULLEC D. ET AL: "The Bisindolylmalemide GF 109203X Is a Potent and Selective Inhibitor of Protein Kinase C", JOURNAL OF BIOLOGICAL CHEMISTRY, vol. 266, no. 24, 1991, pages 15771 - 15781, XP000572210 *

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