Synthesis and bio-molecular study of (+)-N-Acetyl-α-amino acid dehydroabietylamine derivative for the selective therapy of hepatocellular carcinoma.

Muhammad Ayaz Mustufa,Firdous Imran Ali,Imran Ali Hashmi,Imam Bakhsh Solangi,Jameel Ahmed Baig,Naim Ul Hasan Naqvi,Cigdem Ozen,Gokhan Yildiz,Mehmet Ozturk,Shoaib Muhammad,Afshan Aslam

doi:10.1186/s12885-016-2942-5

Abstract

BackgroundThe 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.MethodsAn 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.ResultsHep3B cells were found to be the most sensitive with IC50 of 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.ConclusionIt is concluded that a novel DAAD2 with IC50 values 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.Electronic supplementary materialThe online version of this article (doi:10.1186/s12885-016-2942-5) contains supplementary material, which is available to authorized users.

Highlights

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
Based on primary screening results (Fig. 1 and Additional file 3: Figure S1), DAAD 2 with a functional site of N-acetyl cysteine group at C-18 (2R) was found to be the most potent with 100% growth attenuation at 50 μM and 10 μM concentration on both Huh7 and MCF7 cells
All cells were treated with DAAD2 at different concentrations in triplicates ranging from 10 μM to 0.313 μM; for controls up to

Summary

Introduction

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. Hepatocellular carcinoma (HCC) is the most common primary liver cancer. It is the fifth most frequent neoplasm and the third largest mortality causing cancer [1, 2]. Sorafenib is the first oral multikinase inhibitor and only the approved drug by Food and Drug Administration (FDA) for the treatment of HCC [10, 11]. These disappointing facts and figures suggested a dire need of effective drugs for the treatments of HCC. The (+)-Dehydroabietylamines (DAA) and its derivatives reported for biological activities including cytotoxic effects [12,13,14,15], endo-peptidase inhibitory activity, antibacterial [16, 17], antitumoral [18], and anti-carcinogenesis activity [19]

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