title: Synthesis and bio-molecular study of (+)-N-Acetyl-α-amino acid dehydroabietylamine derivative for the selective therapy of hepatocellular carcinoma.
abstract: The purpose of present work is to synthesize novel (+)-Dehydroabietylamine derivatives (DAAD) using N-acetyl-α-amino acid conjugates and determine its cytotoxic effects on hepatocellular carcinoma cells.
  
  
  
  An analytical study was conducted to explore cytotoxic activity of DAAD on hepatocellular carcinoma cell lines. The cytotoxicity effect was recorded using sulforhodamine B technique. Cell cycle analysis was performed using Propidium Iodide (PI) staining. Based on cell morphology, anti growth activity and microarray findings of DAAD2 treatment, Comet assay, Annexin V/PI staining, Immunoperoxidase assay and western blots were performed accoringly.
  
  
  
  Hep3B cells were found to be the most sensitive with IC50of 2.00 ± 0.4 μM against (+)-N-(N-Acetyl-L-Cysteine)-dehydroabietylamine as DAAD2. In compliance to time dependent morphological changes of low cellular confluence, detachment and rounding of DAAD2 treated cells; noticeable changes in G2/M phase were recorded may be leading to cell cycle cessation. Up-regulation (5folds) of TUBA1A gene in Hep3B cells was determined in microarray experiments. Apoptotic mode of cell death was evaluated using standardized staining procedures including comet assay and annexin V/PI staining, Immuno-peroxidase assay. Using western blotting technique, caspase dependant apoptotic mode of cell death was recorded against Hep3B cell line.
  
  
  
  It is concluded that a novel DAAD2 with IC50values less than 8 μM can induce massive cell attenuation following caspase dependent apoptotic cell death in Hep3B cells. Moreover, the corelation study indicated that DAAD2 may have vital influence on cell prolifration properties.
authors.affiliation: 5th Floor, PHRC Specialized Research Centre on Child Health, National Institute of Child Health, Karachi, 75500, Pakistan. ayazbukero@gmail.com.|Baqai Institute of Pharmaceutical Sciences (BIPS), Baqai Medical University, Karachi, 74600, Pakistan. ayazbukero@gmail.com.|Department of Molecular, Biology and Genetics, BilGen Genetics and Biotechnology Center, Bilkent University, Ankara, 06800, Turkey. ayazbukero@gmail.com.
authors.forename: Muhammad Ayaz
authors.initials: MA
authors.lastname: Mustufa
authors.affiliation: Izmir International Biomedicine and Genome Institute, iBG-izmir, Dokuz Eylül University, 35340, Balcova, Izmir, Turkey.
authors.forename: Cigdem
authors.initials: C
authors.lastname: Ozen
authors.affiliation: Department of Chemistry, University of Karachi, Karachi, 75270, Pakistan.
authors.forename: Imran Ali
authors.initials: IA
authors.lastname: Hashmi
authors.affiliation: Department of Chemistry, University of Karachi, Karachi, 75270, Pakistan.
authors.forename: Afshan
authors.initials: A
authors.lastname: Aslam
authors.affiliation: National Center of Excellence in Analytical Chemistry, University of Sindh, Jamshoro, 76080, Pakistan.
authors.forename: Jameel Ahmed
authors.initials: JA
authors.lastname: Baig
authors.affiliation: Department of Medical Biology, Erzincan University Faculty of Medicine, Erzincan, 24100, Turkey.
authors.forename: Gokhan
authors.initials: G
authors.lastname: Yildiz
authors.affiliation: Department of Chemistry, University of Karachi, Karachi, 75270, Pakistan.
authors.forename: Shoaib
authors.initials: S
authors.lastname: Muhammad
authors.affiliation: Dr. M. A. Kazi Institute of Chemistry, University of Sindh, Jamshoro, 76080, Pakistan.
authors.forename: Imam Bakhsh
authors.initials: IB
authors.lastname: Solangi
authors.affiliation: Baqai Institute of Pharmaceutical Sciences (BIPS), Baqai Medical University, Karachi, 74600, Pakistan.
authors.forename: Naim
authors.initials: N
authors.lastname: Ul Hasan Naqvi
authors.affiliation: Izmir International Biomedicine and Genome Institute, iBG-izmir, Dokuz Eylül University, 35340, Balcova, Izmir, Turkey.
authors.forename: Mehmet
authors.initials: M
authors.lastname: Ozturk
authors.affiliation: Department of Chemistry, University of Karachi, Karachi, 75270, Pakistan. firdousilyas@yahoo.com.
authors.forename: Firdous Imran
authors.initials: FI
authors.lastname: Ali
mesh_terms: D045784:Abietanes / Q000138:chemical synthesis* / Q000494:pharmacology*; D000970:Antineoplastic Agents / Q000138:chemical synthesis* / Q000494:pharmacology*; D017209:Apoptosis / Q000187:drug effects; D006528:Carcinoma, Hepatocellular / Q000235:genetics / Q000378:metabolism; D020169:Caspases / Q000378:metabolism; D002453:Cell Cycle / Q000187:drug effects; D045744:Cell Line, Tumor; D049109:Cell Proliferation / Q000187:drug effects; D059565:G2 Phase Cell Cycle Checkpoints / Q000187:drug effects; D006801:Humans; D020128:Inhibitory Concentration 50; D008113:Liver Neoplasms / Q000235:genetics / Q000378:metabolism
chemical_list: D045784:Abietanes; D000970:Antineoplastic Agents; C000600819:dehydroabietylamine; D020169:Caspases
keywords: Apoptosis; Connectivity map; Cytotoxicity; Dehydroabietylamine derivatives; Hepatocellular carcinoma