Calcium Dibutyryladenosine Cyclophosphate

The aim of the study was to screen new candidate mol.-targeted anti-leukemia compounds with potential functions of targeted up-regulating ID4 gene expression.Promoter region of ID4 gene including the upstream -3000 bp sequence of transcriptional start site and message RNA sequence were fished out.Online promoter anal. tools of TESS and Genomax were used to search possible sequence of transcriptional start site and message RNA sequence were fished out.Online promoter anal. tools of TESS and Genomax were used to search possible cis-acting structure from human transcription factor database.The activity of related drugs with potential effects upon ID4 gene expression was analyzed using SAGE database.GEO database was applied to search the gene expression profiling regulated by ID4 gene.Finally, similar anal. between gene expression profiling by ID4 and genome-wide profiling regulated by 163 known drugs or active compounds was manipulated to screen the drugs and candidate compounds with similar gene expression profiling with ID4 gene.MOLT4 cell line was treated with the above candidate active compounds to investigate the ID4 gene expression by RT-PCR assay.ID4 gene had a type II promoter with a typical TATA box in upstream -45 bp of transcription start site.The 1300 bp-length promoter of ID4 gene contained a few cis-acting structures classified into two function types, i.e. pos. regulatory type, including transcription factors Spl and c-Myb, cAMP, glucocorticoid receptor (GR) and estrogen receptor (ER), and neg. regulatory type, including Wilms tumor-1 (WT1) and early growth response-2 (EGR2).The similarity of gene expression profiling was identified between cAMP and ID4 gene.ID4 gene expression was induced in MOLT4 cell line after treatment with calcium dibutyryladenosine cyclophosphate at the concentration of 0.1 mmol/L.The comprehensive bioinformatic anal., based upon the combination of regulatory sequence prediction of promoter, similarity anal. of gene expression profiling and literature review, could be considered as a practical tool in screening the candidate drugs with the activity of targeted regulating functional genes.Calcium dibutyryladenosine cyclophosphate could induce ID4 gene expression in leukemic cells.

To study the quant. determination of the calcium dibutyryladenosine cyclophosphate and lyophilized powder for injection using HPLC.The content was determined using HPLC.The Recovery rate was 99.4% and RSD was 0.45%.HPLC is fine in precision, highly reproducible, ready to be used, and instant.It may determine the content of calcium dibutyryladenosine cyclophosphate and lyophilized powder for injection with high quality.