Transformation of a Metal Chelate into a “Catch and Anchor” Inhibitor of Botulinum A Protease
Targeting the botulinum neurotoxin light chain (LC) metalloprotease using small-molecule metal chelate inhibitors is a promising approach to counter the effects of the lethal toxin. However, to overcome the pitfalls associated with simple reversible metal chelate inhibitors, it is crucial to investi...
| Main Authors: | , , , , , , , |
|---|---|
| Format: | Article |
| Language: | English |
| Published: |
MDPI AG
2023-02-01
|
| Series: | International Journal of Molecular Sciences |
| Subjects: | |
| Online Access: | https://www.mdpi.com/1422-0067/24/5/4303 |
| _version_ | 1797615284246282240 |
|---|---|
| author | Lucy Lin Ealin N. Patel Alexander L. Nielsen Lewis D. Turner William H. Tepp Kong Nguyen Sabine Pellett Kim Janda |
| author_facet | Lucy Lin Ealin N. Patel Alexander L. Nielsen Lewis D. Turner William H. Tepp Kong Nguyen Sabine Pellett Kim Janda |
| author_sort | Lucy Lin |
| collection | DOAJ |
| description | Targeting the botulinum neurotoxin light chain (LC) metalloprotease using small-molecule metal chelate inhibitors is a promising approach to counter the effects of the lethal toxin. However, to overcome the pitfalls associated with simple reversible metal chelate inhibitors, it is crucial to investigate alternative scaffolds/strategies. In conjunction with Atomwise Inc., in silico and in vitro screenings were conducted, yielding a number of leads, including a novel 9-hydroxy-4<i>H</i>-pyrido [1,2-a]pyrimidin-4-one (PPO) scaffold. From this structure, an additional series of 43 derivatives were synthesized and tested, resulting in a lead candidate with a <i>K</i><sub>i</sub> of 150 nM in a BoNT/A LC enzyme assay and 17 µM in a motor neuron cell-based assay. These data combined with structure-activity relationship (SAR) analysis and docking led to a bifunctional design strategy, which we termed “catch and anchor” for the covalent inhibition of BoNT/A LC. Kinetic evaluation was conducted on structures prepared from this catch and anchor campaign, providing <i>k</i><sub>inact</sub>/<i>K</i><sub>i</sub> values, and rationale for inhibition seen. Covalent modification was validated through additional assays, including an FRET endpoint assay, mass spectrometry, and exhaustive enzyme dialysis. The data presented support the PPO scaffold as a novel candidate for targeted covalent inhibition of BoNT/A LC. |
| first_indexed | 2024-03-11T07:24:08Z |
| format | Article |
| id | doaj.art-ffe576387d604f53aa133ad86b430606 |
| institution | Directory Open Access Journal |
| issn | 1661-6596 1422-0067 |
| language | English |
| last_indexed | 2024-03-11T07:24:08Z |
| publishDate | 2023-02-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | International Journal of Molecular Sciences |
| spelling | doaj.art-ffe576387d604f53aa133ad86b4306062023-11-17T07:46:35ZengMDPI AGInternational Journal of Molecular Sciences1661-65961422-00672023-02-01245430310.3390/ijms24054303Transformation of a Metal Chelate into a “Catch and Anchor” Inhibitor of Botulinum A ProteaseLucy Lin0Ealin N. Patel1Alexander L. Nielsen2Lewis D. Turner3William H. Tepp4Kong Nguyen5Sabine Pellett6Kim Janda7Department of Chemistry and Immunology, The Skaggs Institute for Chemical Biology, Worm Institute of Research and Medicine (WIRM), The Scripps Research Institute, La Jolla, CA 92037, USADepartment of Chemistry and Immunology, The Skaggs Institute for Chemical Biology, Worm Institute of Research and Medicine (WIRM), The Scripps Research Institute, La Jolla, CA 92037, USADepartment of Chemistry and Immunology, The Skaggs Institute for Chemical Biology, Worm Institute of Research and Medicine (WIRM), The Scripps Research Institute, La Jolla, CA 92037, USADepartment of Chemistry and Immunology, The Skaggs Institute for Chemical Biology, Worm Institute of Research and Medicine (WIRM), The Scripps Research Institute, La Jolla, CA 92037, USADepartment of Bacteriology, University of Wisconsin, 1550 Linden Drive, Madison, WI 53706, USAAtomwise Inc., 717 Market Street, Suite 800, San Francisco, CA 94103, USADepartment of Bacteriology, University of Wisconsin, 1550 Linden Drive, Madison, WI 53706, USADepartment of Chemistry and Immunology, The Skaggs Institute for Chemical Biology, Worm Institute of Research and Medicine (WIRM), The Scripps Research Institute, La Jolla, CA 92037, USATargeting the botulinum neurotoxin light chain (LC) metalloprotease using small-molecule metal chelate inhibitors is a promising approach to counter the effects of the lethal toxin. However, to overcome the pitfalls associated with simple reversible metal chelate inhibitors, it is crucial to investigate alternative scaffolds/strategies. In conjunction with Atomwise Inc., in silico and in vitro screenings were conducted, yielding a number of leads, including a novel 9-hydroxy-4<i>H</i>-pyrido [1,2-a]pyrimidin-4-one (PPO) scaffold. From this structure, an additional series of 43 derivatives were synthesized and tested, resulting in a lead candidate with a <i>K</i><sub>i</sub> of 150 nM in a BoNT/A LC enzyme assay and 17 µM in a motor neuron cell-based assay. These data combined with structure-activity relationship (SAR) analysis and docking led to a bifunctional design strategy, which we termed “catch and anchor” for the covalent inhibition of BoNT/A LC. Kinetic evaluation was conducted on structures prepared from this catch and anchor campaign, providing <i>k</i><sub>inact</sub>/<i>K</i><sub>i</sub> values, and rationale for inhibition seen. Covalent modification was validated through additional assays, including an FRET endpoint assay, mass spectrometry, and exhaustive enzyme dialysis. The data presented support the PPO scaffold as a novel candidate for targeted covalent inhibition of BoNT/A LC.https://www.mdpi.com/1422-0067/24/5/4303botulinum neurotoxincatch and anchor inhibitioncovalent inhibitorsstructure-based drug design |
| spellingShingle | Lucy Lin Ealin N. Patel Alexander L. Nielsen Lewis D. Turner William H. Tepp Kong Nguyen Sabine Pellett Kim Janda Transformation of a Metal Chelate into a “Catch and Anchor” Inhibitor of Botulinum A Protease International Journal of Molecular Sciences botulinum neurotoxin catch and anchor inhibition covalent inhibitors structure-based drug design |
| title | Transformation of a Metal Chelate into a “Catch and Anchor” Inhibitor of Botulinum A Protease |
| title_full | Transformation of a Metal Chelate into a “Catch and Anchor” Inhibitor of Botulinum A Protease |
| title_fullStr | Transformation of a Metal Chelate into a “Catch and Anchor” Inhibitor of Botulinum A Protease |
| title_full_unstemmed | Transformation of a Metal Chelate into a “Catch and Anchor” Inhibitor of Botulinum A Protease |
| title_short | Transformation of a Metal Chelate into a “Catch and Anchor” Inhibitor of Botulinum A Protease |
| title_sort | transformation of a metal chelate into a catch and anchor inhibitor of botulinum a protease |
| topic | botulinum neurotoxin catch and anchor inhibition covalent inhibitors structure-based drug design |
| url | https://www.mdpi.com/1422-0067/24/5/4303 |
| work_keys_str_mv | AT lucylin transformationofametalchelateintoacatchandanchorinhibitorofbotulinumaprotease AT ealinnpatel transformationofametalchelateintoacatchandanchorinhibitorofbotulinumaprotease AT alexanderlnielsen transformationofametalchelateintoacatchandanchorinhibitorofbotulinumaprotease AT lewisdturner transformationofametalchelateintoacatchandanchorinhibitorofbotulinumaprotease AT williamhtepp transformationofametalchelateintoacatchandanchorinhibitorofbotulinumaprotease AT kongnguyen transformationofametalchelateintoacatchandanchorinhibitorofbotulinumaprotease AT sabinepellett transformationofametalchelateintoacatchandanchorinhibitorofbotulinumaprotease AT kimjanda transformationofametalchelateintoacatchandanchorinhibitorofbotulinumaprotease |