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Targeted kinases are overexpressed across all three cancer types, and expression levels correlate significantly with overall survival, supporting major roles in cancer development and progression. The work rationalizes synthesis of TAJ4, a site-specific inhibitor, then assesses its cytotoxicity in vitro alongside computational approaches. 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BMC Cancer (2024) 24:1521 [https://doi.org/10.1186/s12885-024-13269-4](https://doi.org/10.1186/s12885-024-13269-4)  \nBMC Cancer  \n RESEARCH Open Access  \nIdentification of dimethyl 2,2’-    \n((methylenebis(2-(2H-benzo[d][1,2,3] triazol-2-yl)-4-(2,4,4-trimethylpentan-2-yl) - 6,1phenylene))bis(oxy))diacetate (TAJ4) as antagonist of NEK-Family: a future for potential drug discovery  \nMubashir Aziz1, Syeda Abida Ejaz1*, Pervaiz Ali Channar2, Ali G. Alkhathami3, Tahir Qadri4, Zahid Hussain4, Mumtaz Hussaain4 and Rabail Ujan5  \nAbstract  \nThe purpose of the current study was to analyze and validate the existing gap in knowledge, by conducting a differential expression analysis and validation of NEK6, NEK7, and NEK9 in breast, cervical, and glioblastoma cancer and targeting these proteins through development of novel site specific inhibitor with favorable pharmacokinetic and safety profile, using open-source databases. The analysis revealed that the targeted kinases were overexpressed in all three types of cancer. Their expression was significantly linked to overall survival rates, which suggests that they playa major role in the development and progression of these cancers. After, having the prognostic importance of These findings provided a rationale for synthesizing novel compound i. e., dimethyl 2,2’-((methylenebis(2-(2H-benzo[d][1,2,3] triazol-2-yl)-4-(2,4,4-trimethylpentan-2-yl)-6,1phenylene))bis(oxy))diacetate (TAJ4)), capable of effectively targeting these proteins using in-vitro cytotoxicity assays and comprehensive computational approaches. Then the inhibitory potential of TAJ4 was evaluated against cell lines of the respective cancers (HeLa cells, MCF-7 cells, and Vero cells) . The growth inhibitory values (GI50) suggested that TAJ4 exhibited strong inhibitory potential towards MCF-7 cells (GI50 = 3.18 ± 0.11 µM) in comparison to the HeLa cell line (GI50 = 8.12 ± 0.43 µM), surpassing that of standard drugs. Furthermore, in-silico investigations, including density functional theory (DFT) calculations and molecular docking studies, revealed a substantial reactivity profile of TAJ4, with promising molecular interactions against NEK7, NEK9, TP53, NF-KAPPA-B, and caspase-3 proteins. Further investigation using in-vitro and in-vivo approaches is recommended to fully establish the therapeutic efficacy and safety profile of TAJ4 .  \nKeywords NIMA related kinases, Expression analysis, Benzotriazole derivative, Single-crystal analysis, Cytotoxicity assay, In-silico studies  \n*Correspondence: Syeda Abida Ejaz [abida.ejaz@iub.edu.pk](abida.ejaz@iub.edu.pk)  \nFull list of author information is available at the end of the article  \n© The Author(s) 2024. Open Access This article is licensed under a Creative Commons Attribution-NonCommercial-NoDerivatives 4.0 International License, which permits any non-commercial use, sharing, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons licence, and indicate if you modified the licensed material. You do not have permission under this licence to share adapted material derived from this article or parts of it. The images or other third party material in this article are included in the article’s Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons licence and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this licence, visit [http://creativecommons.org/licenses/by-nc-nd/4.0/](http://creativecommons.org/licenses/by-nc-nd/4.0/.)[.](http://creativecommons.org/licenses/by-nc-nd/4.0/.)  \nAziz et al. BMC Cancer (2024) 24:1521  \nIntroduction  \nCancer is still one of the main causes of death in the world. Breast, cervical, and glioblastoma cancers con","cbCaitCpdHlziUmY","https://ap.wps.com/l/cbCaitCpdHlziUmY","pdf",6125279,22,"English","# Abstract\n# Introduction","[{\"question\":\"Which NEK family members are investigated and in which cancer types?\",\"answer\":\"The study focuses on NEK6, NEK7, and NEK9 in breast cancer, cervical cancer, and glioblastoma, using open-source databases for differential expression and validation.\"},{\"question\":\"What is the main biological finding from the expression and survival analysis?\",\"answer\":\"The targeted kinases are overexpressed in all three cancer types, and their expression is significantly linked to overall survival, indicating involvement in development and progression.\"},{\"question\":\"How was TAJ4 evaluated and what cytotoxicity result was highlighted?\",\"answer\":\"TAJ4 was assessed using in-vitro cytotoxicity assays and computational approaches. GI50 values indicate stronger inhibition against MCF-7 cells (3.18 ± 0.11 µM) than HeLa cells (8.12 ± 0.43 µM).\"}]","Identification of dimethyl 2,2’-((methylenebis(2-(2H-benzo[d][1,2,3] triazol-2-yl)-4-(2,4,4-trimethylpentan-2-yl)-6,1phenylene))bis(oxy))diacetate (TAJ4) as antagonist of NEK-Family - a future for potential drug discovery | PDF",1789985037,55]