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Updated: Aug 5, 2026
Kinase Inhibitor Screening In Self-assembled Human Protein Microarrays
Published on: October 23, 2019
Targeting Human Protein Kinase CK2 by a Library of Indeno[1,2-b]Indoles: Contribution of Thermal Shift Assay to
Matheus M Guimarães1,2, Christian Werner3, Belen Leroy1
1Gastroenterology and Technologies for Health Team, Centre de Recherche en Cancérologie de Lyon, Centre Léon Bérard, CNRS 5286, INSERM 1052, Université Claude Bernard Lyon 1, Univ. Lyon, Lyon, France.
Abstract:
Protein kinase CK2 is the subject of numerous studies in medicinal chemistry due to its involvement in the development of several diseases, primarily cancers. Its overexpression in tumor cells is related to key processes such as tumor immune evasion and cell proliferation. The scientific approach of this study aims to investigate the thermal shift assay (TSA) as a pre-screening tool and to complement it with a co-crystallization approach in post-screening. Therefore, the synthesis of seven small-molecule CK2 inhibitors derived from indeno[1,2-b]indoles was supplemented by 18 related derivatives from our in-house compound library. The 25 molecules belong to four sub-scaffolds, namely 4b,9b-dihydroxy-4b,5,6,7,8,9b-hexahydroindeno[1,2-b]indole-9,10-dione (D-0), 5,6,7,8-tetrahydroindeno[1,2-b]indole-9,10-dione (D-1), 9-hydroxy-5H-indeno[1,2-b]indol-10-one (D-2), and 5H-indeno[1,2-b]indole-6,9,10-trione (D-3). The most active CK2 inhibitors identified by capillary electrophoresis (CE)-based assay belong to the D-1 sub-scaffold. In the TSA, these compounds also generate significant shifts of the melting temperature (Tm) of CK2, indicating a clear correlation between the results of the CE-based assay and those of the TSA. The contribution of co-crystallization in post-screening also demonstrated the effectiveness of D-1 sub-scaffold compared with D-0 sub-scaffold.
Insights
This study evaluates thermal shift assay (TSA) and co-crystallization as methods for identifying protein kinase CK2 inhibitors. The D-1 sub-scaffold compounds showed the most promise, correlating well between assays.
Area of Science:
- Medicinal Chemistry
- Biochemistry
- Structural Biology
Background:
- Protein kinase CK2 is implicated in cancer development, influencing tumor immune evasion and cell proliferation.
- Targeting CK2 is a key strategy in cancer therapy.
- Developing effective CK2 inhibitors requires robust screening methods.
Purpose of the Study:
- To investigate the efficacy of thermal shift assay (TSA) as a pre-screening tool for CK2 inhibitors.
- To complement pre-screening with co-crystallization for post-screening validation.
- To identify potent CK2 inhibitors from indeno[1,2-b]indole derivatives.
Main Methods:
- Synthesis of 25 small-molecule CK2 inhibitors across four sub-scaffolds (D-0, D-1, D-2, D-3).
- Screening using capillary electrophoresis (CE)-based assays.
- Thermal Shift Assay (TSA) to measure changes in CK2 melting temperature (Tm).
- Co-crystallization studies for structural validation.
Main Results:
- The D-1 sub-scaffold compounds exhibited the highest activity in the CE-based assay.
- TSA results showed a strong correlation with the CE-based assay, with active compounds causing significant Tm shifts.
- Co-crystallization confirmed the effectiveness of the D-1 sub-scaffold over the D-0 sub-scaffold.
Conclusions:
- TSA is a reliable pre-screening tool for identifying CK2 inhibitors.
- The D-1 sub-scaffold represents a promising chemical class for developing novel CK2 inhibitors.
- Combined CE, TSA, and co-crystallization approaches provide a comprehensive strategy for inhibitor discovery.
