1. A continuous casting method for manufacturing thin cast strips, in which a thin cast strip is cast while molten metal is being supplied into a molten metal pool formed by a pair of cooling drums and a pair of side dams, scum dams dipped into molten metal of the molten metal pool, and the thin cast strip is wound into a coil so as to manufacture a plurality of coil-shaped thin cast strips, the continuous casting method for manufacturing thin cast strips comprising the steps of pulling up the scum dams from the molten metal and blowing gas onto a molten metal face in the molten metal pool toward one of the pair of cooling drums or both of the pair of cooling drums when the casting work, corresponding to one coil of the thin cast strip, is completed.
2. A continuous casting machine for manufacturing thin cast strips, in which a thin cast strip is cast while molten metal is being supplied into a molten metal pool formed by a pair of cooling drums and a pair of side dams, and the thin cast strip is wound into a coil so as to manufacture a plurality of coil-shaped thin cast strips, the continuous casting machine for manufacturing thin cast strips comprising gas nozzles to blow gas, which are arranged at an upper central portion of the molten metal pool, being directed toward one or both of the pair of cooling drums, to a molten metal face of the molten metal pool when casting operation corresponding to one coil of the thin cast strip has been completed, and a pair of scum dams arranged on both sides of a molten metal pouring nozzle, said scum dams are moveable into and out of the molten metal pool.
The claims below are in addition to those above.
All refrences to claim(s) which appear below refer to the numbering after this setence.
1. A mu-conotoxin peptide essentially comprising the amino acid sequence:
SEQ\u2003ID\u2003NO:\u20031
Xaa1-Xaa2-Cys-Cys-Xaa3-Xaa4-Xaa5-Xaa6-Xaa7-Cys-
Xaa8-Xaa9-Xaa10-Xaa11-Cys-Xaa12-Xaa13-Xaa14-
Xaa15-Xaa16-Cys-Cys-Xaa17,
a biologically active fragment thereof, a salt thereof, a combination of biologically active fragments thereof, a combination of salts thereof, 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 hydroxylamino acid,
Xaa9 is any non-aromatic hydroxylamino 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 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 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 is absent.
5. The mu-conotoxin peptide of claim 1, wherein
the N-modification of amino-acid Xaa1 is selected from the group consisting of acetylation, formylation, myristoylation and amidation;
Xaa3 and Xaa13 are independently selected from the group consisting of 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 consisting of arginine (Arg), lysine (Lys) and histidine (His);
Xaa8 and Xaa9 are independently selected from the group consisting of serine (Ser) and threonine (Thr);
Xaa11 is selected from the group consisting of phenylalanine (Phe), tyrosine (Tyr), and tryptophan (Trp);
Xaa14 is selected from the group consisting of arginine (Arg), lysine (Lys), histidine (His), cysteine (Cys) and methionine (Met);
Xaa15 is selected from the group consisting of glycine (Gly), alanine (Ala), valine (Val), leucine (Leu), isoleucine (Ile), serine (Ser), threonine (Thr), methionine (Met), cysteine (Cys) and proline (Pro);
Xaa17 is selected from the group consisting of 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
SEQ\u2003ID\u2003NO:\u20032
pGlu-Gly-Cys-Cys-Asn-Gly-Pro-Lys-Gly-Cys-Ser-Ser-
Lys-Trp-Cys-Arg-Asp-His-Ala-Arg-Cys-Cys,
a biologically active fragment thereof, a salt thereof, a combination of biologically active fragments thereof, a combination of salts thereof, 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) andor 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 andor adjuvants.
10. A method of treating pain, the method comprising administering the pharmaceutical composition of claim 9 to a subject in need thereof.
11. The method of claim 9, wherein the pain is migraine, acute pain, persistent pain, chronic pain, neuropathic pain or nociceptive pain.
12. A method for the treatment of a disorder associated with voltage-sensitive sodium channels in a subject in need thereof, 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 in need thereof, 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 the patient 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 claim 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 to a subject in need thereof.
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 in a subject in need thereof, the method comprising administering a pharmaceutically effective amount of at least one mu-conotoxin peptide of claim 1 to the subject.
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, or buccal routes of application.
25. The anesthetic of claim 24, wherein it is in the form of tablets, capsules, lozenges, dental pastes, suppositories, inhalants, solutions, ointments, creams or 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. An implantable device comprising the pharmaceutical composition of claim 9.