Inhibition of the Clathrin Terminal Domain—Amphiphysin Protein–Protein Interaction. Probing the Pitstop 2 Aromatic Moiety
Fuente:
Wiley Open Access
Saved in:
| Main Authors: | , , , , , , , , , , |
|---|---|
| Format: | Artículo Open Access |
| Published: |
Wiley
2025
|
| Subjects: | |
| Online Access: | |
| Tags: |
Add Tag
No Tags, Be the first to tag this record!
|
| _version_ | 1867008816595337216 |
|---|---|
| author | Kate Prichard Mark J. Robertson Ngoc Chau Jing Xue Martin Neuenschwander Uwe Fink Andre Horatscheck Marc Nazare Phillip J. Robinson Volker Haucke Adam McCluskey |
| author_facet | Kate Prichard Mark J. Robertson Ngoc Chau Jing Xue Martin Neuenschwander Uwe Fink Andre Horatscheck Marc Nazare Phillip J. Robinson Volker Haucke Adam McCluskey Kate Prichard Mark J. Robertson Ngoc Chau Jing Xue Martin Neuenschwander Uwe Fink Andre Horatscheck Marc Nazare Phillip J. Robinson Volker Haucke Adam McCluskey |
| collection | Wiley Open Access |
| contents | Inhibition of the Clathrin Terminal Domain—Amphiphysin Protein–Protein Interaction. Probing the Pitstop 2 Aromatic Moiety Kate Prichard Mark J. Robertson Ngoc Chau Jing Xue Martin Neuenschwander Uwe Fink Andre Horatscheck Marc Nazare Phillip J. Robinson Volker Haucke Adam McCluskey ChemMedChem Pitstop 2, (Z)‐N‐(5‐(4‐bromenzylidene)‐4‐oxo‐4,5‐dihydrothiazol‐2‐yl)naphthalene‐1‐sulfonamide (1) inhibits the clathrin terminal domain–amphiphysin interaction (NTD‐PPI) and has been widely used to investigate endocytosis. Herein, the synthesis of 56 novel Pitstop 2 analogues via four discrete focused libraries is reported. Specific modification to the 4‐bromonenzylidene moiety is explored, and their ability to inhibit the NTD‐PPI interaction is examined by ELISA. In cell endocytosis, effects are measured for selective analogues as is the inhibition of dynamin, another protein involved in the endocytosis process. The most NTD‐PPI active analogues retain 2‐ and 4‐disposed substituents on the aromatic head, with 2,3,4‐trihydroxy (28) the most active (IC50 0.94 μm). Catechol‐free 2,3‐dihydroxybenzo[b][1,4]dioxone (54) is a more promising lead with an NTD‐PPI IC50 = 1.16 μm. The corresponding benzo[d][1,3]dioxole (53) is threefold less active suggesting ring size preference at this position. Nine analogues show improved or comparable NTD‐PPI activity to Pitstop 2 with IC50 values from 0.94 to 2.1 μM. Heterocyclic analogues are well tolerated and potent inhibitors of CME in U2OS cells, in particular, benzofuran 67 (NTD‐PPI IC50 1.5 μm and CME IC50 6.8 ± 2.7 μm). This positions 67 as one of the most cell active inhibitors of clathrin‐mediated endocytosis yet reported. 10.1002/cmdc.202500321 http://creativecommons.org/licenses/by/4.0/ |
| doi_str_mv | 10.1002/cmdc.202500321 |
| format | Artículo Open Access |
| id | wiley_oa_10_1002_cmdc_202500321 |
| institution | Wiley Open Access |
| license_str_mv | http://creativecommons.org/licenses/by/4.0/ |
| publishDate | 2025 |
| publisher | Wiley |
| record_format | wiley_oa |
| spellingShingle | Inhibition of the Clathrin Terminal Domain—Amphiphysin Protein–Protein Interaction. Probing the Pitstop 2 Aromatic Moiety Kate Prichard Mark J. Robertson Ngoc Chau Jing Xue Martin Neuenschwander Uwe Fink Andre Horatscheck Marc Nazare Phillip J. Robinson Volker Haucke Adam McCluskey ChemMedChem Inhibition of the Clathrin Terminal Domain—Amphiphysin Protein–Protein Interaction. Probing the Pitstop 2 Aromatic Moiety Kate Prichard Mark J. Robertson Ngoc Chau Jing Xue Martin Neuenschwander Uwe Fink Andre Horatscheck Marc Nazare Phillip J. Robinson Volker Haucke Adam McCluskey ChemMedChem Pitstop 2, (Z)‐N‐(5‐(4‐bromenzylidene)‐4‐oxo‐4,5‐dihydrothiazol‐2‐yl)naphthalene‐1‐sulfonamide (1) inhibits the clathrin terminal domain–amphiphysin interaction (NTD‐PPI) and has been widely used to investigate endocytosis. Herein, the synthesis of 56 novel Pitstop 2 analogues via four discrete focused libraries is reported. Specific modification to the 4‐bromonenzylidene moiety is explored, and their ability to inhibit the NTD‐PPI interaction is examined by ELISA. In cell endocytosis, effects are measured for selective analogues as is the inhibition of dynamin, another protein involved in the endocytosis process. The most NTD‐PPI active analogues retain 2‐ and 4‐disposed substituents on the aromatic head, with 2,3,4‐trihydroxy (28) the most active (IC50 0.94 μm). Catechol‐free 2,3‐dihydroxybenzo[b][1,4]dioxone (54) is a more promising lead with an NTD‐PPI IC50 = 1.16 μm. The corresponding benzo[d][1,3]dioxole (53) is threefold less active suggesting ring size preference at this position. Nine analogues show improved or comparable NTD‐PPI activity to Pitstop 2 with IC50 values from 0.94 to 2.1 μM. Heterocyclic analogues are well tolerated and potent inhibitors of CME in U2OS cells, in particular, benzofuran 67 (NTD‐PPI IC50 1.5 μm and CME IC50 6.8 ± 2.7 μm). This positions 67 as one of the most cell active inhibitors of clathrin‐mediated endocytosis yet reported. 10.1002/cmdc.202500321 http://creativecommons.org/licenses/by/4.0/ |
| title | Inhibition of the Clathrin Terminal Domain—Amphiphysin Protein–Protein Interaction. Probing the Pitstop 2 Aromatic Moiety |
| topic | ChemMedChem |
| url | https://chemistry-europe.onlinelibrary.wiley.com/doi/10.1002/cmdc.202500321 |