Methods for treating pain and screening analgesic compounds
The present invention relates to the use of compounds that block the α9α10 subtype of the nicotinic acetylcholine receptor (nAChR) for treating pain, such as neuropathic pain and inflammatory pain, and inflammatory disorders, such as arthritis. The present invention also relates to screening compounds to identify analgesic agents that block the α9α10 subtype of the nAChR.
1. A method for treating or preventing conditions or disorders associated with the α9α10 subtype of the nicotinic acetylcholine receptor (nAChR) in an individual which comprises administering to an individual in need thereof a therapeutically effective amount of an active agent or a pharmaceutically acceptable salt thereof, wherein the active agent blocks the α9α10 subtype of the nAChR, wherein the active agent is selected from the group consisting of: α-conotoxin PeIA (SEQ ID NO:3),
an analog of α-conotoxin Vc1.1 (SEQ ID NO:2), the analog of Vc1.1 being at least one selected from the group consisting of Glu14iodo-His Vc1.1; Glu14iodo-Tyr Vc1.1; Glu14iodo-Trp Vc1.1; Glu14iodo-Phe Vc1.1; an addition to the C-terminus of the preceding analogs; an addition to the N-terminus of the preceding analogs; additions to the C-terminus and N-terminus of the preceding analogs; and replacement of Gly1 in Vc1.1; and
an analog of α-conotoxin PeIA (SEQ ID NO:3), the analog of PeIA being at least one selected from the group consisting of Glu14iodo-His PeIA; Glu14iodo-Tyr PeIA; Glu14iodo-Trp PeIA; Glu14iodo-Phe PeIA; an addition to the C-terminus of PeIA or the preceding analogs; an addition to the N-terminus of PeIA or the preceding analogs; additions to the C-terminus and N-terminus of PeIA or the preceding analogs; and replacement of Gly1 in PeIA.
2. The method of claim 1 , wherein the condition is pain and the administration of the active agent alleviates pain in the individual.
3. The method of claim 1 , wherein the condition is inflammation mediated by immune cells and the administration of the active agent reduces inflammation.
4. The method of claim 3 , wherein the inflammation is associated with rheumatic diseases.
5. A method of inhibiting migration of immune cells in an individual in need thereof which comprises administering to an individual an immune cell migration-inhibiting amount of an active agent or a pharmaceutically acceptable salt thereof, wherein said active agent blocks the α9α10 subtype of the nicotinic acetylcholine receptor (nAChR), wherein the active agent is selected from the group consisting of: α-conotoxin PeIA (SEQ ID NO:3),
an analog of α-conotoxin Vc1.1 (SEQ ID NO:2), the analog of Vc1.1 being at least one selected from the group consisting of Glu14iodo-His Vc1.1; Glu14iodo-Tyr Vc1.1; Glu14iodo-Trp Vc1.1; Glu14iodo-Phe Vc1.1; an addition to the C-terminus of the preceding analogs; an addition to the N-terminus of the preceding analogs; additions to the C-terminus and N-terminus of the preceding analogs; and replacement of Gly1 in Vc1.1; and
an analog of α-conotoxin PeIA (SEQ ID NO:3), the analog of PeIA being at least one selected from the group consisting of Glu14iodo-His PeIA; Glu14iodo-Tyr PeIA; Glu14iodo-Trp PeIA; Glu14iodo-Phe PeIA; an addition to the C-terminus of PeIA or the preceding analogs; an addition to the N-terminus of PeIA or the preceding analogs; additions to the C-terminus and N-terminus of PeIA or the preceding analogs; and replacement of Gly1 in PeIA.
6. A method for identifying drug candidates for use as treating or preventing conditions or disorders associated with the α9α10 subtype of the nicotinic acetylcholine receptor (nAChR) or for inhibiting the migration of immune cells comprising:
screening a drug candidate agent for its ability to block the activity of the α9α10 subtype of the nAChR, wherein screening the drug candidate agent comprises displacing a labeled α-conotoxin from the α9α10 subtype of the nAChR by a candidate drug agent and measuring the labeled α-conotoxin, wherein a decrease in labeled α-conotoxin indicates displacement of the labeled α-conotoxin from the α9α10 subtype of the nAChR, and identifies the candidate drug agent as a drug candidate,
wherein the α-conotoxin is selected from the group consisting of:
an analog of α-conotoxin RgIA (SEQ ID NO:1), the analog of RgIA being at least one selected from the group consisting of Arg9citrulline RgIA; Arg9ω-nitro-Arg RgIA; Tyr10iodo-Tyr RgIA; Tyr10Trp RgIA; Tyr10Phe RgIA; Arg9citrulline, Tyr10iodo-Tyr RgIA; Arg9ω-nitro-Arg, Tyr10iodo-Tyr RgIA; Ser4Ala RgIA; RgIA-Cys-amide; an addition to the C-terminus of RgIA or the preceding analogs; an addition to the N-terminus of the preceding analogs; additions to the C-terminus and N-terminus of RgIA or the preceding analogs; replacement of Arg13 in RgIA, and replacement of Gly1 in RgIA;
an analog of α-conotoxin Vc1.1 (SEQ ID NO:2), the analog of Vc1.1 being at least one selected from the group consisting of Glu14iodo-His Vc1.1; Glu14iodo-Tyr Vc1.1; Glu14iodo-Trp Vc1.1; Glu14iodo-Phe Vc1.1; an addition to the C-terminus of Vc1.1 or the preceding analogs; an addition to the N-terminus of Vc1.1 or the preceding analogs; additions to the C-terminus and N-terminus of Vc1.1 or the preceding analogs; and replacement of Gly1 in Vc1.1; and
an analog of α-conotoxin PeIA (SEQ ID NO:3), the analog of PeIA being at least one selected from the group consisting of Glu14iodo-His PeIA; Glu14iodo-Tyr PeIA; Glu14iodo-Trp PeIA; Glu14iodo-Phe PeIA; an addition to the C-terminus of PeIA or the preceding analogs; an addition to the N-terminus of PeIA or the preceding analogs; additions to the C-terminus and N-terminus of PeIA or the preceding analogs; and replacement of Gly1 in PeIA.
7. A method for identifying drug candidates for use in treating or preventing conditions or disorder associated with the α9α10 subtype of the nicotinic acetylcholine receptor (nAChR) or for inhibiting the migration of immune cells, the method comprising carrying out an in vitro or in vivo biological assay on a drug candidate and carrying out the same assay on an α-conotoxin, wherein the results obtained from the biological assay on the drug candidate and the α-conotoxin are compared, wherein the α-conotoxin is selected from the group consisting of:
an analog of α-conotoxin RgIA (SEQ ID NO:1), the analog of RgIA being at least one selected from the group consisting of Arg9citrulline RgIA; Arg9ω-nitro-Arg RgIA; Tyr10iodo-Tyr RgIA; Tyr10Trp RgIA; Tyr10Phe RgIA; Arg9citrulline, Tyr10iodo-Tyr RgIA; Arg9ω-nitro-Arg, Tyr10iodo-Tyr RgIA; Ser4Ala RgIA, RgIA-Cys-amide; an addition to the C-terminus of RgIA or the preceding analogs; an addition to the N-terminus of the preceding analogs; additions to the C-terminus and N-terminus of RgIA or the preceding analogs; replacement of Arg13 in RgIA and replacement of Gly1 in RgIA;
an analog of α-conotoxin Vc1.1 (SEQ ID NO:2), the analog of Vc1.1 being at least one selected from the group consisting of Glu14iodo-His Vc1.1; Glu14iodo-Tyr Vc1.1; Glu14iodo-Trp Vc1.1; Glu14iodo-Phe Vc1.1; an addition to the C-terminus of Vc1.1 or the preceding analogs; addition to the N-terminus of Vc1.1 or the preceding analogs; additions to the C-terminus and N-terminus of Vc1.1 or the preceding analogs; and replacement of Gly1 in Vc1.1; and
an analog of α-conotoxin PeIA (SEQ ID NO:3), the analog of PeIA being at least one selected from the group consisting of Glu14iodo-His PeIA; Glu14iodo-Tyr PeIA; Glu14iodo-Trp PeIA; Glu14iodo Phe PeIA; an addition to the C-terminus of PeIA or the preceding analogs; an addition to the N-terminus of PeIA or the preceding analogs; additions to the C-terminus and N-terminus of PeIA or the preceding analogs; and replacement of Gly1 in PeIA.
8. A method for treating or preventing conditions or disorders associated with the α9α10 subtype of the nicotinic acetylcholine receptor (nAChR) in an individual which comprises administering to an individual in need thereof a therapeutically effective amount of an active agent or a pharmaceutically acceptable salt thereof, wherein the active agent blocks the α9α10 subtype of the nAChR, wherein the active agent is selected from the group consisting of:
an analog of α-conotoxin RgIA (SEQ ID NO:1), the analog of RgIA being at least one selected from the group consisting of Arg9citrulline RgIA; Arg9ω-nitro-Arg RgIA; Tyr10iodo-Tyr RgIA; Tyr10Trp RgIA; Tyr10Phe RgIA; Arg9citrulline, Tyr10iodo-Tyr RgIA; Arg9ω-nitro-Arg, Tyr10iodo-Tyr RgIA; Ser4Ala RgIA; RgIA-Cys-amide; an addition to the C-terminus of RgIA or the preceding analogs; an addition to the N-terminus of the preceding analogs; additions to the C-terminus and N-terminus of RgIA or the preceding analogs; replacement of Arg13 in RgIA, and replacement of Gly1 in RgIA.
9. The method of claim 8 , wherein the condition is pain and the administration of the active agent alleviates pain in the individual.
10. The method of claim 8 , wherein the condition is inflammation mediated by immune cells and the administration of the active agent reduces inflammation.
11. The method of claim 10 , wherein the inflammation is associated with rheumatic diseases.
12. A method of inhibiting migration of immune cells in an individual in need thereof which comprises administering to an individual an immune cell migration-inhibiting amount of an active agent or a pharmaceutically acceptable salt thereof, wherein said active agent blocks the α9α10 subtype of the nicotinic acetylcholine receptor (nAChR), wherein the active agent is selected from the group consisting of:
an analog of α-conotoxin RgIA (SEQ ID NO:1), the analog of RgIA being at least one selected from the group consisting of Arg9citrulline RgIA; Arg9ω-nitro-Arg RgIA; Tyr10iodo-Tyr RgIA; Tyr10Trp RgIA; Tyr10Phe RgIA; Arg9citrulline, Tyr10iodo-Tyr RgIA; Arg9ω-nitro-Arg, Tyr10iodo-Tyr RgIA; Ser4Ala RgIA; RgIA-Cys-amide; an addition to the C-terminus of RgIA or the preceding analogs; an addition to the N-terminus of RgIA or the preceding analogs; additions to the C-terminus and N-terminus of RgIA or the preceding analogs; replacement of Arg13 in RgIA, and replacement of Gly1 in RgIA.