Triple-negative breast cancer(TNBC)is a highly aggressive and metastasizing cancer that has the worst prognosis out of all breast cancer subtypes.The epithelial emesenchymal transition(EMT)and cancer stem cells(CSCs)h...Triple-negative breast cancer(TNBC)is a highly aggressive and metastasizing cancer that has the worst prognosis out of all breast cancer subtypes.The epithelial emesenchymal transition(EMT)and cancer stem cells(CSCs)have been proposed as important mechanisms underlying TNBC metastasis.CDK9 is highly expressed in breast cancer,including TNBC,where it promotes EMT and induces cancer cell stemness.In this study,we have identified a tetrahydroisoquinoline derivative(compound 1)as a potent and selective CDK9-cyclin T1 inhibitor via virtual screening.Interestingly,by targeting the ATP binding site,compound 1 not only inhibited CDK9 activity but also disrupted the CDK9-cyclin T1 proteineprotein interaction(PPI).Mechanistically,compound 1 reversed EMT and reduced the ratio of CSCs by blocking the CDK9-cyclin T1 interaction,leading to reduced TNBC cell proliferation and migration.To date,compound 1 is the first reported tetrahydroisoquinoline-based CDK9-cyclin T1 ATP-competitive inhibitor that also interferes with the interaction between CDK9 and cyclin T1.Compound 1 may serve as a promising scaffold for developing more selective and potent anti-TNBC agents.Our work also provides insight into the role of the CDK9-cyclin T1 PPI on EMT and CSCs and highlights the feasibility and significance of targeting CDK9 for the treatment of TNBC.展开更多
Objective： To investigate the effect of berberine, one of the main alkaloids of Rhizoma coptidis, on myocardial morphology and the expressions of cyclin-dependent kinase 9 （Cdk9） and cyclin T1 protein in the myocar...Objective： To investigate the effect of berberine, one of the main alkaloids of Rhizoma coptidis, on myocardial morphology and the expressions of cyclin-dependent kinase 9 （Cdk9） and cyclin T1 protein in the myocardium of type 2 diabetic rats. Methods： Type 2 diabetes mellitus rats were induced by an injection of 35 mg/kg streptozotocin （STZ） and a high-carbohydrate/high-fat diet for 16 weeks. Diabetic rats were given low-, middle-, high-dose berberine （75, 150, 300 mg/kg）, fenofibrate （100 mg/kg） and rosiglitazone （4 mg/kg） for another 16 weeks, respectively. The myocardium structure was observed with hematoxylin ＆ eosin （H＆E） staining and Cdk9 and cyclin T1 protein expressions were detected by immunohistochemistry. Results： Middle-dose, high-dose berberine improved myocardial hypertrophy and interstitial fibrosis of diabetic rats. Cdk9 and cyclin T1 protein were significantly lower in diabetic myocardium than in control one （P〈0.01）, and middle-dose, high-dose berberine and fenofibrate obviously increased both Cdk9 and cyclin T1 expression to near control level （P〈0.01）. Conclusion： Berberine modulates Cdk9 and cyclin T1 protein expression in diabetic myocardium which may contribute to ameliorate myocardium damage.展开更多
The aberrant overexpression of cyclin-dependent kinase 9 (CDK9) in cancer cells results in the loss of proliferative control, making it an attractive therapeutic target for various cancers. However, the highly structu...The aberrant overexpression of cyclin-dependent kinase 9 (CDK9) in cancer cells results in the loss of proliferative control, making it an attractive therapeutic target for various cancers. However, the highly structural similarity between CDK9 and CDK2 makes the development of novel selective CDK9 inhibitors a challenging task and thus limits their clinical applications. Here, an effective two-stage virtual screening strategy was developed to identify novel CDK9 inhibitors with better inhibitory activity and higher selectivity. The first screening stage aims to select potential compounds with better inhibitory activity than Roniciclib, one of the most effective CDK9 inhibitors, through reliable structure-based pharmacophoric virtual screening and accurate molecular docking analyses. The second stage employs a very detailed visual inspection process, in which several structural criteria describing the major difference between the binding pockets of CDK9 and CDK2 are taken into consideration, to identify compounds with higher selectivity than CAN508, one of the CDK9 inhibitors with distinguished selectivity. Finally, three compounds (NCI207113 from NCI database and TCM0004 and TCM3282 from TCM database) with better inhibitory activity and higher selectivity were successfully identified as novel CDK9 inhibitors. These three compounds also display excellent binding stabilities, great pharmacokinetic properties and low toxicity in MD simulations and ADMET predictions. Besides, the results of binding free energy calculations suggest that enhancing van der Waals interaction and nonpolar solvation energy and/or reducing polar solvation energy can significantly improve the binding affinity of these CDK9 inhibitors. Their clinical potentials to serve as anticancer drug candidates can be further evaluated through a series of <em>in vitro/in vivo</em> bioassays in the future. To the best of our knowledge, this is the first attempt to identify novel CDK9 inhibitors with both better inhibitory activity and higher selectivity through an effective two-stage virtual screening strategy.展开更多
基金This work was supported by the Health and Medical Research Fund(No.HMRF/14150561)the National Natural Science Foundation of China(No.201575121 and 21775131)+3 种基金the Hong Kong Baptist University Century Club Sponsorship Scheme 2020,Teaching Development Fund(No.TDG/1920/02)the Science and Technology Development Fund,Macao SAR,China(File no.0072/2018/A2 and 0007/2020/A1)SKLQRCM(UM)-2020-2022the University of Macao,China(MYRG2019e00002eICMS).
文摘Triple-negative breast cancer(TNBC)is a highly aggressive and metastasizing cancer that has the worst prognosis out of all breast cancer subtypes.The epithelial emesenchymal transition(EMT)and cancer stem cells(CSCs)have been proposed as important mechanisms underlying TNBC metastasis.CDK9 is highly expressed in breast cancer,including TNBC,where it promotes EMT and induces cancer cell stemness.In this study,we have identified a tetrahydroisoquinoline derivative(compound 1)as a potent and selective CDK9-cyclin T1 inhibitor via virtual screening.Interestingly,by targeting the ATP binding site,compound 1 not only inhibited CDK9 activity but also disrupted the CDK9-cyclin T1 proteineprotein interaction(PPI).Mechanistically,compound 1 reversed EMT and reduced the ratio of CSCs by blocking the CDK9-cyclin T1 interaction,leading to reduced TNBC cell proliferation and migration.To date,compound 1 is the first reported tetrahydroisoquinoline-based CDK9-cyclin T1 ATP-competitive inhibitor that also interferes with the interaction between CDK9 and cyclin T1.Compound 1 may serve as a promising scaffold for developing more selective and potent anti-TNBC agents.Our work also provides insight into the role of the CDK9-cyclin T1 PPI on EMT and CSCs and highlights the feasibility and significance of targeting CDK9 for the treatment of TNBC.
文摘Objective： To investigate the effect of berberine, one of the main alkaloids of Rhizoma coptidis, on myocardial morphology and the expressions of cyclin-dependent kinase 9 （Cdk9） and cyclin T1 protein in the myocardium of type 2 diabetic rats. Methods： Type 2 diabetes mellitus rats were induced by an injection of 35 mg/kg streptozotocin （STZ） and a high-carbohydrate/high-fat diet for 16 weeks. Diabetic rats were given low-, middle-, high-dose berberine （75, 150, 300 mg/kg）, fenofibrate （100 mg/kg） and rosiglitazone （4 mg/kg） for another 16 weeks, respectively. The myocardium structure was observed with hematoxylin ＆ eosin （H＆E） staining and Cdk9 and cyclin T1 protein expressions were detected by immunohistochemistry. Results： Middle-dose, high-dose berberine improved myocardial hypertrophy and interstitial fibrosis of diabetic rats. Cdk9 and cyclin T1 protein were significantly lower in diabetic myocardium than in control one （P〈0.01）, and middle-dose, high-dose berberine and fenofibrate obviously increased both Cdk9 and cyclin T1 expression to near control level （P〈0.01）. Conclusion： Berberine modulates Cdk9 and cyclin T1 protein expression in diabetic myocardium which may contribute to ameliorate myocardium damage.
文摘The aberrant overexpression of cyclin-dependent kinase 9 (CDK9) in cancer cells results in the loss of proliferative control, making it an attractive therapeutic target for various cancers. However, the highly structural similarity between CDK9 and CDK2 makes the development of novel selective CDK9 inhibitors a challenging task and thus limits their clinical applications. Here, an effective two-stage virtual screening strategy was developed to identify novel CDK9 inhibitors with better inhibitory activity and higher selectivity. The first screening stage aims to select potential compounds with better inhibitory activity than Roniciclib, one of the most effective CDK9 inhibitors, through reliable structure-based pharmacophoric virtual screening and accurate molecular docking analyses. The second stage employs a very detailed visual inspection process, in which several structural criteria describing the major difference between the binding pockets of CDK9 and CDK2 are taken into consideration, to identify compounds with higher selectivity than CAN508, one of the CDK9 inhibitors with distinguished selectivity. Finally, three compounds (NCI207113 from NCI database and TCM0004 and TCM3282 from TCM database) with better inhibitory activity and higher selectivity were successfully identified as novel CDK9 inhibitors. These three compounds also display excellent binding stabilities, great pharmacokinetic properties and low toxicity in MD simulations and ADMET predictions. Besides, the results of binding free energy calculations suggest that enhancing van der Waals interaction and nonpolar solvation energy and/or reducing polar solvation energy can significantly improve the binding affinity of these CDK9 inhibitors. Their clinical potentials to serve as anticancer drug candidates can be further evaluated through a series of <em>in vitro/in vivo</em> bioassays in the future. To the best of our knowledge, this is the first attempt to identify novel CDK9 inhibitors with both better inhibitory activity and higher selectivity through an effective two-stage virtual screening strategy.
