Expanding the small-molecule toolbox for studying CLC chloride channel physiology and biophysics
Abstract/Contents
- Abstract
- The chloride channel (CLC) family is a class of membrane proteins that regulates the flux of chloride ions across cell membranes. These ion channels play critical roles in electrical excitation of muscles and neurons, as well as in maintaining proper water and salt balance in the body. CLCs are traditionally difficult drug targets, as demonstrated by the low potency and general lack of homolog selectivity among existing classes of CLC inhibitors. To address this challenge, we employed a rational-design approach to develop a novel, small-molecule scaffold that displays unprecedented selectivity for one CLC homolog. Specifically, this molecule inhibits one of the two homologs expressed in the kidney, CLC-Ka, with > 20-fold higher potency than the next most closely related CLC homolog, which is 90% identical in sequence. Following a similar approach, we identified and optimized a new class of potent and selective inhibitors for CLC-2, the most abundant voltage-gated chloride channel expressed in the brain. Data from genetic knockout studies in mice suggest a critical role for CLC-2 in ion homeostasis and electrical excitability in the central nervous system (CNS), but the mechanisms for these processes have been difficult to elucidate in the absence of specific and potent chemical modulators of CLC-2. Structure-activity relationship studies have allowed us to develop these inhibitors into a best-in-class molecule that represents the first and only nanomolar potency small-molecule inhibitor to target the CLC family. In addition to extraordinary selectivity among CLCs, we demonstrated that this molecule is highly selective for CLC-2 within the context of the brain, showing no off-target effects among a diverse panel of CNS receptors, channels, and transporters that we screened. Additionally, this inhibitor acutely and specifically blocks CLC-2 current in wild-type hippocampal CA1 neurons with no effect on CLC-2--/-- neurons. This exciting new molecular tool acts as a fast, reversible, and specific inhibitor of CLC-2 function in the brain and should enable study of CLC-2 in the CNS.
Description
| Type of resource | text |
|---|---|
| Form | electronic resource; remote; computer; online resource |
| Extent | 1 online resource. |
| Place | California |
| Place | [Stanford, California] |
| Publisher | [Stanford University] |
| Copyright date | 2019; ©2019 |
| Publication date | 2019; 2019 |
| Issuance | monographic |
| Language | English |
Creators/Contributors
| Author | Koster, Anna Katherine |
|---|---|
| Degree supervisor | Du Bois, Justin |
| Degree supervisor | Maduke, Merritt C, 1966- |
| Thesis advisor | Du Bois, Justin |
| Thesis advisor | Maduke, Merritt C, 1966- |
| Thesis advisor | Cegelski, Lynette |
| Degree committee member | Cegelski, Lynette |
| Associated with | Stanford University, Department of Chemistry. |
Subjects
| Genre | Theses |
|---|---|
| Genre | Text |
Bibliographic information
| Statement of responsibility | Anna Katherine Koster. |
|---|---|
| Note | Submitted to the Department of Chemistry. |
| Thesis | Thesis Ph.D. Stanford University 2019. |
| Location | electronic resource |
Access conditions
- Copyright
- © 2019 by Anna Katherine Koster
- License
- This work is licensed under a Creative Commons Attribution Non Commercial 3.0 Unported license (CC BY-NC).
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