Human voltage gated sodium channel beta1A subunit and methods of use
AssigneeOrtho-McNeil Pharmaceutical, Inc.
InventorsNing Qin, Ellen Codd, Michael D'Andrea
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Abstract
DNAs encoding human voltage gated sodium channel β1A subunit have been cloned and characterized. The recombinant protein is capable of forming biologically active protein. The cDNA's have been expressed in recombinant host cells that produce active recombinant protein. The recombinant protein is also purified from the recombinant host cells. In addition, the recombinant host cells are utilized to establish a method for identifying modulators of the receptor activity, and receptor modulators are identified.
Claims34
An isolated polynucleotide that encodes a human β1A sodium channel subunit protein, said polynucleotide comprising a member selected from a group consisting of:
- (a) a polynucleotide having at least a 75% identity to a polynucleotide encoding a polypeptide consisting of amino acids 1 to 268 of SEQ.ID.NO.:14;
- (b) a polynucleotide having at least 75% identity to a polynucleotide encoding a polypeptide consisting of amino acids 150 to 268 of SEQ.ID.NO.:14;
- (c) a polynucleotide which is complementary to the polynucleotide of (a) or (b);
- and
- (d) a polynucleotide comprising at least 15 sequential bases of the polynucleotide of (a), (b), or (c).
An expression vector for expression of a human β1A sodium channel subunit protein in a recombinant host, wherein said vector contains a recombinant gene encoding a human β1A sodium channel subunit protein and functional derivatives thereof.
A recombinant host cell containing a recombinantly cloned gene encoding Human β1A sodium channel subunit protein or a functional derivative thereof.
An isolated protein encoded by a nucleic acid sequence capable of hybridizing under stringent hybridization conditions to a nucleotide sequence having the sequence of SEQ ID NO:12 or SEQ ID NO:13 that when combined with a Human α sodium channel subunit protein in a cell permits sodium ion flux in the cell.
A monospecific antibody immunologically reactive with a human β1A sodium channel subunit protein.
A process for expression of a Human β1A sodium channel subunit protein in a recombinant host cell, comprising:
- (a) introducing an expression vector comprising a nucleic acid sequence capable of hybridizing under stringent hybridization conditions to a nucleotide sequence, or its complementary sequence, having the sequence of SEQ ID NO:12 or SEQ ID NO:13 into a cell;
- (b) culturing the cell of step (a) under conditions which allow expression of a a protein encoded by the nucleotide seuqence.
A method of screening for a modulator of sodium channel activity comprising:
- (a) providing a cell that co-expresses a protein encoded by a nucleic acid capable of hybridizing under stringent hybridization conditions to a nucleotide sequence, or its complementary sequence, represented by SEQ ID NO:12 or SEQ ID NO:13 and a sodium channel α subunit protein wherein the cell elicits a sodium ion flux;
- (b) contacting the cell with a putative β1A modulating compound;
- and
- (c) measuring a change upon the cell that alters the sodium ion flux.
A method of treating neuropathic pain in a patient in need of such treatment comprising altering the level of a human β1A subunit in a dorsal root ganglia cell in the patient.
A pharmaceutical composition comprising a compound of claim useful for use as a local anesthetic.
A method for decreasing neuropathic pain in an individual comprising administering to said individual a modulator of a sodium channel β1A subunit in an amount effective to change the sodium channel activity in said individual.
A method for treating neuropathic pain in a subject comprising altering the level of sodium channel β1A subunits on the surface of a cell in a subject.
A method for decreasing neuropathic pain in a human comprising the step of administering a sodium channel β1A subunit-binding molecule to a sodium channel β1A subunit-expressing cell in the human.
Dependencies are inferred from the claim text.