title: Biological evaluation of 2-arylidene-4, 7-dimethyl indan-1-one (FXY-1): a novel Akt inhibitor with potent activity in lung cancer.
abstract: OBJECTIVE
  Human lung cancer is contributed to be a major mortality factor in the cancer-related deaths. Arylidene indan-ones constitute a new class of potential anti-tumor compounds. Herein, we report the biological efficacy of the 2-arylidene-4, 7-dimethyl indan-1-one (FXY-1), a potential lead molecule of arylidene indan-ones in lung cancer models. We previously described anticancer activity of FXY-1 against human breast adenocarcinoma.
  
  
  METHODS
  FXY-1 efficacy was assessed by standard anticancer screening in NCI-H 460, A549, NCI-H 1975 and NCI-H 2170 cells. Initial molecular docking analysis was performed to check the interaction of compound to Akt enzyme. Anti-proliferation, anti-metastatic and transendothelial cell migration were performed to check efficacy of the drug. Western blot analysis was performed to understand the regulation of pro-apoptotic and anti-apoptotic proteins by the compound. The effect of FXY-1 on caspase induction and Akt phosphorylation were checked using Western blot analysis. Flow cytometry was used to reveal the cell cycle changes and apoptosis-inducing properties of FXY-1 in the lung cancer cells. In-vitro Akt inhibition property of the compound was studied using a fluorescence-based, coupled-enzyme reaction. The in-vivo efficacy of the compound was determined using a mouse xenograft model.
  
  
  RESULTS
  Our molecular docking analysis reveals higher interaction of FXY-1 with Akt. FXY-1 depicted anti-proliferative and pro-apoptotic activity with higher therapeutic window in-vitro in NCI-H 460 and A549 cells. The compound treatment to lung cancer cells resulted in induction of DNA fragmentation, inhibition of transendothelial migration, caspase activation and poly (ADP-ribose) polymerase (PARP) cleavage. FXY-1 treatment resulted in G 0/G 1 arrest in both cell lines at lower concentrations, but led to apoptosis at higher doses. Western blot analysis revealed dephosphorylation of Akt (Ser 473) with activation of p53, Bax, Bak, Bid and reduction in Bcl-2 and Bcl-xL levels. Further mechanistic investigation showed that FXY-1 activity was facilitated through an allosteric inhibition of Akt enzyme. FXY-1 treatment significantly reduced the tumor growth and pAkt levels in mouse xenograft exhibiting the in-vivo efficacy in the compound.
  
  
  CONCLUSIONS
  Collectively, our results suggest DNA damage-mediated activation by FXY-1 in lung cancer cells leading to extensive apoptosis through the mitochondrial pathway.
authors.affiliation: Department of Clinical Laboratory Sciences, College of Applied Medical Sciences, King Khalid University, P.O. Box # 641, Abha, Saudi Arabia. prachu.rg@gmail.com.
authors.forename: Prasanna
authors.initials: P
authors.lastname: Rajagopalan
authors.affiliation: Department of Medical Rehabilitation Sciences, College of Applied Medical Sciences, King Khalid University, P.O. Box # 641, Abha, Saudi Arabia.
authors.forename: Khalid A
authors.initials: KA
authors.lastname: Alahmari
authors.affiliation: Department of Biochemistry, Faculty of Science, Alexandria University, Alexandria, Egypt.
authors.forename: Ashraf A
authors.initials: AA
authors.lastname: Elbessoumy
authors.affiliation: Department of Geriatrics, Bioinformatics Program, University of Arkansas for Medical Sciences and University of Arkansas at Little Rock, Little Rock, AR, 72205, USA.
authors.forename: Meenakshisundaram
authors.initials: M
authors.lastname: Balasubramaniam
authors.affiliation: Department of Medicinal chemistry, Krisani Bioscience Pvt LTD, Hyderabad, India.
authors.forename: Radhakrishnan
authors.initials: R
authors.lastname: Suresh
authors.affiliation: Department of Physiology, College of Medicine, King Khalid University, P.O.Box # 641, Abha, Saudi Arabia.
authors.forename: Mohammed Eajaz Ahmed
authors.initials: ME
authors.lastname: Shariff
authors.affiliation: Department of Microbiology and Parasitology, College of Medicine, King Khalid University, P.O.Box # 641, Abha, Saudi Arabia.
authors.forename: Harish C
authors.initials: HC
authors.lastname: Chandramoorthy
mesh_terms: D000818:Animals; D000970:Antineoplastic Agents / Q000494:pharmacology*; D017209:Apoptosis / Q000187:drug effects*; D051017:Apoptosis Regulatory Proteins / Q000378:metabolism; D001597:Benzylidene Compounds / Q000494:pharmacology*; D002453:Cell Cycle / Q000187:drug effects; D045744:Cell Line, Tumor; D049109:Cell Proliferation / Q000187:drug effects; D006801:Humans; D007189:Indans / Q000494:pharmacology*; D008175:Lung Neoplasms / Q000188:drug therapy* / Q000378:metabolism; D051379:Mice; D062105:Molecular Docking Simulation; D047428:Protein Kinase Inhibitors / Q000494:pharmacology*; D051057:Proto-Oncogene Proteins c-akt / Q000037:antagonists & inhibitors*; D023041:Xenograft Model Antitumor Assays
chemical_list: C000608566:2-benzylidene-4,7-dimethylindan-1-one; D000970:Antineoplastic Agents; D051017:Apoptosis Regulatory Proteins; D001597:Benzylidene Compounds; D007189:Indans; D047428:Protein Kinase Inhibitors; D051057:Proto-Oncogene Proteins c-akt
keywords: Akt; Apoptosis; Arylidene; Cell cycle; FXY-1; Indan-one; Lung cancer