Mu-Conotoxin Peptides and Use Thereof as a Local Anesthetic

Abstract

In one embodiment, the present invention relates to novel mu-conotoxin peptides, biologically active fragments thereof, combinations thereof and/or variants thereof. An embodiment of the invention also relates to their use in pharmaceutical composition for the treatment or prevention of pain, and their use in the preparation of an anesthetic.

1 . A mu-conotoxin peptide comprising the amino acid sequence:
 Xaa1-Xaa2-Cys-Cys-Xaa3-Xaa4-Xaa5-Xaa6-Xaa7-Cys-Xaa8-Xaa9-Xaa10-Xaa11-Cys-Xaa12-Xaa13-Xaa14-Xaa15-Xaa16-Cys-Cys-Xaa17 [SEQ ID No 1]   a biologically active fragment thereof, a salt thereof, a combination thereof and/or variants thereof, and wherein   Xaa1 is any N-modified amino acid,   Xaa2 is glycine,   Xaa3 is any acidic amino acid or any of its amide form,   Xaa4 is glycine,   Xaa5 is proline or hydroxy-proline,   Xaa6 is any basic amino acid,   Xaa7 is glycine,   Xaa8 is any non-aromatic hydroxyl amino acid,   Xaa9 is any non-aromatic hydroxyl amino acid,   Xaa10 is any basic amino acid,   Xaa11 is any aromatic amino acid,   Xaa12 is any basic amino acid,   Xaa13 is any acidic amino acid or any of its amide form,   Xaa14 is any basic amino acid, or any sulfur-containing amino acid,   Xaa15 is any hydrophobic or apolar amino acid, or any non-aromatic hydroxyl amino acid,   Xaa16 is any basic amino acid,   Xaa17 is absent or is any apolar amino acid, or an amide group.   
     
     
         2 . The mu-conotoxin peptide of  claim 1 , wherein at least one amino acid consisting of amino acids Xaa3, Xaa4, Xaa5, Xaa6 and Xaa7, or any combination thereof, is absent. 
     
     
         3 . The mu-conotoxin peptide of  claim 1 , wherein at least one amino acid consisting of amino acids Xaa8, Xaa9, Xaa10 and Xaa11, or any combination thereof, is absent. 
     
     
         4 . The mu-conotoxin peptide of  claim 1 , wherein at least one amino acid consisting of amino acids Xaa12, Xaa13, Xaa14, Xaa15 and Xaa16, or any combination thereof, is absent. 
     
     
         5 . The mu-conotoxin peptide of  claim 1 , wherein
 the N-modification of amino-acid Xaa1 is selected from the group comprising acetylation, formylation, myristoylation or amidation;   Xaa3 and Xaa13 are independently selected from the group comprising aspartic acid (Asp), asparagine (Asn), glutamic acid (Glu), glutamine (Gln) and pyroglutamic acid (pGlu or Z);   Xaa6, Xaa10, Xaa12 and Xaa16 are independently selected from the group comprising arginine (Arg), lysine (Lys) and histidine (His);   Xaa8 and Xaa9 are independently selected from the group comprising serine (Ser) and threonine (Thr);   Xaa11 is selected from the group comprising phenylalanine (Phe), tyrosine (Tyr), and tryptophane (Trp);   Xaa14 is selected from the group comprising arginine (Arg), lysine (Lys) and histidine (His), cysteine (Cys) and methionine (Met);   Xaa15 is selected from the group comprising glycine (Gly), alanine (Ala), valine (Val), leucine (Leu) and isoleucine (Ile), serine (Ser), threonine (Thr), methionine (Met), cystein (Cys) and proline (Pro);   Xaa17 is selected from the group comprising glycine (Gly), alanine (Ala), valine (Val), leucine (Leu), isoleucine (Ile), threonine (Thr), methionine (Met), phenylalanine (Phe) and proline (Pro).   
     
     
         6 . The mu-conotoxin peptide of  claim 1 , wherein Xaa1 is pyroglutamate (pGlu). 
     
     
         7 . The mu-conotoxin peptide of  claim 1 , wherein the amino acid sequence is
 pGlu-Gly-Cys-Cys-Asn-Gly-Pro-Lys-Gly-Cys-Ser-Ser-Lys-Trp-Cys-Arg-Asp-His-Ala-Arg-Cys-Cys [SEQ ID No 2],   a biologically active fragment thereof, a salt thereof, a combination thereof and/or variants thereof.   
     
     
         8 . An isolated and purified nucleic acid sequence comprising
 i) a nucleotide sequence encoding a mu-conotoxin peptide of  claim 1 ,   ii) a nucleic acid sequence complementary to i),   iii) a degenerated nucleic acid sequence of i) or ii),   iv) a nucleic acid sequence capable of hybridizing under stringent conditions to i), ii) or iii),   v) a nucleic acid sequence encoding a truncation or an analog of the mu-conotoxin peptide of  claim 1 ,   vi) and/or a fragment of i), ii), iii), iv) or v) encoding a biologically active fragment of said mu-conotoxin peptide of  claim 1 .   
     
     
         9 . A pharmaceutical composition comprising as an active substance a pharmaceutically effective amount of at least one mu-conotoxin peptide of  claim 1 , optionally in combination with pharmaceutically acceptable carriers, diluents and/or adjuvants. 
     
     
         10 . The pharmaceutical composition of  claim 9  for the treatment or prevention of a pain. 
     
     
         11 . The pharmaceutical composition of  claim 10 , wherein the pain is migraine, acute pain, persistent pain, chronic pain, neuropathic pain or nociceptive pain. 
     
     
         12 . A method for the treatment or prevention of a disorder associated with voltage-sensitive sodium channels in a subject, the method comprising administering a pharmaceutically effective amount of the pharmaceutical composition of  claim 9  to the subject. 
     
     
         13 . The method according to  claim 12 , wherein the voltage-sensitive sodium channels are Nav1.4 channels. 
     
     
         14 . A method for providing anesthesia to a subject, the method comprising administering a pharmaceutically effective amount of the pharmaceutical composition of  claim 9  to the subject. 
     
     
         15 . A method for providing musculoskeletal relaxation in a patient undergoing a surgical procedure requiring anesthesia which comprises administering to a patient in need thereof a pharmaceutically effective amount of at least one mu-conotoxin peptide of  claim 1 . 
     
     
         16 . The method of  claim 15 , wherein the at least one mu-conotoxin peptide is administered as an ocular anesthetic. 
     
     
         17 . The method of  claim 15 , wherein the at least one mu-conotoxin peptide is administered as a local anesthetic. 
     
     
         18 . The method of  claims 15  having an anesthetic effect comprised between about 30 min to 48 hours. 
     
     
         19 . A method for local anesthesia, the method comprising administering a pharmaceutically effective amount of at least one mu-conotoxin peptide of  claim 1 . 
     
     
         20 . The method for local anesthesia of  claim 19  having an anesthetic effect comprised between about 30 min to 48 hours. 
     
     
         21 . A method for the treatment or prevention of a pain, the method comprising administering a pharmaceutically effective amount of at least one mu-conotoxin of  claim 1 . 
     
     
         22 . The method of  claim 21  having an effect comprised between about 30 min to 12 hours. 
     
     
         23 . An anesthetic comprising the pharmaceutical composition of  claim 9 . 
     
     
         24 . The anesthetic of  claim 23 , wherein said anesthetic is suitable for subcutaneous, intravenous, intradermal, intramuscular, intraperitoneal, intranasal, transdermal, buccal routes application. 
     
     
         25 . The anesthetic of  claim 24 , wherein it is in the form of tablets, capsules, lozenges, dental pastes, suppositories, inhalants, solutions, ointments, creams and parenteral depots. 
     
     
         26 . The anesthetic of  claim 25 , wherein the inhalant is a spray. 
     
     
         27 . A vector comprising an isolated and purified nucleic acid sequence of  claim 8 . 
     
     
         28 . A implantable device comprising the pharmaceutical composition of  claim 9 . 
     
     
         29 . A method for providing musculoskeletal relaxation in a patient undergoing a surgical procedure requiring anesthesia which comprises administering to a patient in need thereof a pharmaceutically effective amount of the pharmaceutical composition of  claim 9 . 
     
     
         30 . The method of  claim 29 , wherein the at least one mu-conotoxin peptide is administered as an ocular anesthetic. 
     
     
         31 . The method of  claim 29 , wherein the at least one mu-conotoxin peptide is administered as a local anesthetic. 
     
     
         32 . The method of  claims 29  having an anesthetic effect comprised between about 30 min to 48 hours. 
     
     
         33 . A method for local anesthesia, the method comprising administering a pharmaceutically effective amount of the pharmaceutical composition of  claim 9 . 
     
     
         34 . The method for local anesthesia of  claim 33  having an anesthetic effect comprised between about 30 min to 48 hours. 
     
     
         35 . A method for the treatment or prevention of a pain, the method comprising administering a pharmaceutically effective amount of the pharmaceutical composition of  claim 9 . 
     
     
         36 . The method of  claim 35  having an effect comprised between about 30 min to 12 hours. 
     
     
         37 . An anesthetic comprising the mu-conotoxin peptide of  claim 1 . 
     
     
         38 . The anesthetic of  claim 37 , wherein said anesthetic is suitable for subcutaneous, intravenous, intradermal, intramuscular, intraperitoneal, intranasal, transdermal, buccal routes application. 
     
     
         39 . The anesthetic of  claim 38 , wherein it is in the form of tablets, capsules, lozenges, dental pastes, suppositories, inhalants, solutions, ointments, creams and parenteral depots. 
     
     
         40 . The anesthetic of  claim 39 , wherein the inhalant is a spray. 
     
     
         41 . A implantable device comprising the mu-conotoxin peptide of  claim 1 .