Modified ligand-gated ion channels and methods of use
AssigneeHoward Hughes Medical Institute
InventorsScott Sternson, Peter Lee, Christopher Magnus
Life of this patent
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Abstract
This document relates to materials and methods for modulating ligand gated ion channel (LGIC) activity. For example, modified LGICs including at least one LGIC subunit having a modified ligand binding domain (LBD) and/or a modified ion pore domain (IPD) are provided. Also provided are exogenous LGIC ligands that can bind to and activate the modified LGIC, as well as methods of modulating ion transport across the membrane of a cell of a mammal, methods of modulating the excitability of a cell in a mammal, and methods of treating a mammal having a channelopathy.
Claims25
(canceled)
A synthetic nucleic acid construct encoding a modified ligand gated ion channel (LGIC) subunit comprising:
- (i) a human alpha 7 nicotinic acetylcholine receptor (α7-nAChR) ligand binding domain (LBD) comprising an L131G amino acid substitution, a Q139L amino acid substitution, and/or a Y217F amino acid substitution as numbered in SEQ ID NO:2;
- and
- (ii) an ion pore domain (IPD);
- wherein an exogenous LGIC ligand selected from the group consisting of compound 0780, compound 0782, compound 0785, compound 0788, compound 0782, compound 0789, compound 0791, compound 0793, compound 0794, compound 0795, compound 0798, compound 0799, compound 0800, compound 0801, compound 0802, compound 0803, compound 0804, compound 0805, compound 0807, compound 0808, compound 0812, compound 0813, compound 815, compound 816, compound 817, compound 0783, compound 0784, compound 0790, and compound 0792 selectively binds the modified LGIC subunit.
A synthetic nucleic acid construct encoding a modified ligand gated ion channel (LGIC) subunit comprising:
- (i) a human alpha 7 nicotinic acetylcholine receptor (α7-nAChR) ligand binding domain (LBD) having an amino acid substitution at one or more amino acid residues selected from the group consisting of residues 77, 79, 115, 131, 139, 141, 175, 210, 216, 217, and 219 as numbered in SEQ ID NO:2;
- (ii) a 5HT3 ion pore domain (IPD);
- wherein said synthetic nucleic acid construct comprises a nucleic acid sequence having at least 75 percent sequence identity to the sequence set forth in SEQ ID NO:27;
- wherein an exogenous LGIC ligand selected from the group consisting of compound 0780, compound 0782, compound 0785, compound 0788, compound 0782, compound 0789, compound 0791, compound 0793, compound 0794, compound 0795, compound 0798, compound 0799, compound 0800, compound 0801, compound 0802, compound 0803, compound 0804, compound 0805, compound 0807, compound 0808, compound 0812, compound 0813, compound 815, compound 816, compound 817, compound 0783, compound 0784, compound 0790, and compound 0792 selectively binds the modified LGIC subunit.
A synthetic nucleic acid construct encoding a modified ligand gated ion channel (LGIC) subunit comprising:
- (i) a human alpha 7 nicotinic acetylcholine receptor (α7-nAChR) ligand binding domain (LBD) having an amino acid substitution at one or more amino acid residues selected from the group consisting of residues 77, 79, 115, 131, 139, 141, 175, 210, 216, 217, and 219 as numbered in SEQ ID NO:2;
- (ii) a GlyR ion pore domain (IPD);
- and
- wherein said synthetic nucleic acid construct comprises a nucleic acid sequence having at least 75 percent sequence identity to the sequence set forth in SEQ ID NO:28;
- wherein an exogenous LGIC ligand selected from the group consisting of compound 0780, compound 0782, compound 0785, compound 0788, compound 0782, compound 0789, compound 0791, compound 0793, compound 0794, compound 0795, compound 0798, compound 0799, compound 0800, compound 0801, compound 0802, compound 0803, compound 0804, compound 0805, compound 0807, compound 0808, compound 0812, compound 0813, compound 815, compound 816, compound 817, compound 0783, compound 0784, compound 0790, and compound 0792 selectively binds the modified LGIC subunit.
A synthetic nucleic acid construct encoding a modified ligand gated ion channel (LGIC) subunit comprising:
- (i) a human alpha 7 nicotinic acetylcholine receptor (α7-nAChR) ligand binding domain (LBD) having an amino acid substitution at one or more amino acid residues selected from the group consisting of residues 77, 79, 115, 131, 139, 141, 175, 210, 216, 217, and 219 as numbered in SEQ ID NO:2;
- (ii) a GABA ion pore domain (IPD);
- and
- wherein said synthetic nucleic acid construct comprises a nucleic acid sequence having at least 75 percent sequence identity to the sequence set forth in SEQ ID NO:29;
- wherein an exogenous LGIC ligand selected from the group consisting of compound 0780, compound 0782, compound 0785, compound 0788, compound 0782, compound 0789, compound 0791, compound 0793, compound 0794, compound 0795, compound 0798, compound 0799, compound 0800, compound 0801, compound 0802, compound 0803, compound 0804, compound 0805, compound 0807, compound 0808, compound 0812, compound 0813, compound 815, compound 816, compound 817, compound 0783, compound 0784, compound 0790, and compound 0792 selectively binds the modified LGIC subunit.
Dependencies are inferred from the claim text.