国家食药监管总局发布《生物制品稳定性研究技术指导原则》.pdf

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1、中国医药生物技术2015年6月第1O卷第3期ChinMedBiotechnol，June2015，Vo1．10，No．3241Identificationandanti—osteoprosisactivityresearchofup—regulatorsofbonemorphogeneticprotein-2LIXue—hong，GONGShi—qiang，HANXiao—wan，SIShu-yi，WANGYan—hong[Abstract]objectiveTodiscoverpotentialsmall—molecularcompoundswiththeanti—osteoporo

2、sisfunctionsbyup—regulatingtheexpressionofbonemorphogeneticprotein-2(BMP-2)．MethodsTheprocessofscreeningwasperformedusingacell·basedhigh--throughputscreeningmodelforBMP--2withgenisteinasapositivecontro1．Runx2isamajorosteogenictranscriptionfactorinBMP一2一mediatedpathway,andSmadl／5／8isaveryimportan

3、tsignalingmoleculeinthispathway．ThemRNAandproteinexpressionlevelsofBMP一2，Runx2andSmadl／5／8phosphorylationweremeasuredbyrealtimePCRandWesternblot，respectively．TheactivityofALPwasmeasuredbythemethodofPNPEResultsCompoundE19773withtheactivityofup—regulatingtheexpressionofBMP一2wasfoundbyPMBmodels，and

4、theEC5ovalueofE19773was46．2gmol／L．E19773wasfoundtosignificantlyincreasethemRNAexpressionlevelsofBMP一2andRunx2，aswellasincreasetheproteinexpressionlevelsofBMP-2，Runx2andSmadl／5／8phosphorylationinU一2OScells．／nvitroresultsofALPactivityindicatedthatE19773wasabletopromoteosteoblastdiferentiation．Conc

5、lusionCompoundofE19773canup—regulatetheexpressionofBMP一2throughBMP—Smadpathwayattheconcentrationof0．75—50gmol／L．ThusthecompoundCanberegardedasaleadcompoundforthedevelopmentofnewdrugstopreventandtreatosteoporosisinthefuture．[Keywords]Osteoporosis；High—throughputscreening；Bonemorphogeneticprotein2

6、AuthorAfiliations：CollegeofPharmacy,HeilongjiangUniversityofChineseMedicine，Harbin150040，China(LIXue—hong，WANGYan—hong)；InstituteofMedicinalBiotechnology,ChineseAcademyofMedicalSciences&PekingUnionMedicalCollege．Beijing100050，China(LIXue—hong，GONGShi-qiang，HANXiao—wan，SIShu—yi)；InstituteofBasicM

7、edicalSciences，ChineseAcademyofMediealSciences&PekingUnionMedicalCollege，Beijing100005，China(GONGShi—qiang)CorrespondingAuthors：SIShu—yi，Email：sisyimb@hotmail．corn；WANGYan—hong，Email：wang．yanhong@163．cornwwc．netcChinMedBiote