Pharmacodynamic Evaluation And Mechanism Research Of MDMX Transcription Inhibitor In Treating Malignant Glioma

Background and purpose:Glioblastoma(GBM)is a subtype of glioma with the highest degree of malignancy.Its incidence rate accounts for about 40% of the primary intracranial tumors and is increasing year by year.It has the characteristics of high mortality and poor prognosis,which seriously threatens human health.The existing treatment methods for GBM are limited to surgery and chemotherapy,making it difficult to cure.The discovery of specific targets and the development of active molecules are crucial for the development of targeted GBM drugs.Murine Double Minute X(MDMX)is an important factor driving tumor malignancy and is generally highly expressed in GBM cells.The potential anti-tumor activity of chemically targeted MDMX in gliomas is not yet clear.Nicotinamide N-methyltransferase(NNMT)is a key factor that regulates gene expression in tumor cells.It is also highly expressed in GBM cells and promotes their malignancy,making it a potential target for clinical diagnosis and treatment of GBM.However,the relevant mechanism of action has not yet been elucidated.In the early stage of this study,it was found that MDMX transcription inhibitor XI-011 activates p53 activity by inhibiting MDMX transcription activity,thereby exerting anti-tumor activity.This provides a theoretical basis for the development of new and effective anti malignant glioma drugs using MDMX as a therapeutic target.Method and results:1.Evaluation of the activity of XI-011 against malignant glioma cellsThis study synthesized compound XI-011.Through MTT experiments,cell growth curve experiments,colony formation experiments,and Western blot experiments,it was confirmed that XI-011 can inhibit the proliferation of malignant glioma cells,inhibit the expression of MDMX protein in GBM cells,and activate the expression of wild-type p53 protein.2.Evaluation of in vivo anti malignant glioma cell activity of XI-011Healthy C57BL/6 mice were given 10 mg/kg of XI-011 by intraperitoneal injection.Blood and tissue samples were collected at different time points.LC-MS detection showed that XI-011 could quickly distribute in the abdominal cavity to plasma and various tissues,indicating that compound XI-011 could effectively penetrate the blood-brain barrier.BALB/c nude mice were randomly divided into control group,Temozolomide group(50 mg/kg),XI-011 group(10 mg/kg),XI-011 group and Temozolomide group.The experimental results showed that XI-011 alone could effectively inhibit the growth of malignant glioma cells in vivo,and its anti-tumor activity could be enhanced when combined with Temozolomide.3.Study on the related mechanisms of XI-011 in treating malignant gliomaIn this study,NNMT was identified as an upstream factor of MDMX protein and promoted the expression of MDMX protein in GBM cells through molecular docking,bio touch interference technology,gene silencing technology,q RT-PCR,WB and double Luciferase reporter gene technology.Through Pull down,gene silencing,WB,q RT-PCR,double Luciferase reporter gene and technology,we found and identified the transcription factor MYH9 that NNMT regulates the expression of MDMX protein.The mechanism study shows that NNMT plays an anti-malignant role by indirectly regulating the expression of MDMX protein through transcription factors.Research conclusion:This study synthesized compound XI-011,and pharmacodynamic evaluation found that compound XI-011 has good anti-malignant glioma activity in vitro and in vivo;Pharmacokinetic studies have found that compound XI-011 can effectively penetrate the blood-brain barrier;Mechanism research has found that compound XI-011 binds to NNMT protein and indirectly affects the transcription of MDMX through transcription factors,thereby exerting anti malignant glioma effects.