Cellular internalization, cytotoxicity and DNA binding property of 2,3-diaminophenazine, synthesized using Jeanbandyite, a heterogeneous catalyst

Abstract

In recent years, the development of novel chemistry routes for the synthesis of organic compounds has attracted special attention. 2,3-Diaminophenazine (DAP), a derivative of Phenazine, is a large group of nitrogen-containing heterocyclic compound with diverse chemical structure and various biological activities, such as antibacterial, antimicrobial, anti-inflammatory, and anticancer activities. Phenazine is a fluorescent molecule with wide range of biological properties. Therefore, a novel chemical methodology is required for effective synthesis of this product. Numerous oxidants can easily oxidize ortho-phenylenediamine (OPD) to create luminous DAP. This article discusses a simple, sustainable, and safe way to synthesize DAP using water as a green solvent and Jeanbandyite as a catalyst. Mass spectrometry, ¹H-NMR and ¹³C-NMR were used to characterize the molecule, and the catalytic efficacy of Jeanbandyite was assessed. The cellular uptake and cytotoxicity of DAP were investigated to determine whether DAP can be used as a bioprobe in bioapplications. Finally, DAP binding to DNA was methodically performed and confirmed using molecular docking.

Keywords: Catalysis; Cellular internalization; Green chemistry; Jeanbandyite; Phenazine.

Fig. 1

¹H-NMR and ¹³C-NMR spectra (inset) of DAP.

Fig. 2

Mass spectra of DAP.

Scheme 1

(a) Synthetic route and (b) Plausible mechanism for DAP formation.

Fig. 3

Catalytic recyclability studies (% of yield vs. cycle number).

Fig. 4

Cellular internalization of DAP in HeLa cells. The green fluorescence was emitted from DAP, and blue from Hoechst33342. (a) Incubation time of 30 min (b) 1 h (scale bar: 10 µm); Cytotoxicity measurement by MTT assay. (c) HeLa and (d) HEY A8 cells were treated with DAP for 24 h; (e) Fluorescence emission spectra of DAP (0.002 M) in the presence of increasing [DNA] (1.24 × 10^–6–1.15 × 10^–5 M). (f) Change in color of DAP in presence and absence of DNA.

Fig. 5

(a) Ground State optimized structure of DAP (b) The Orientations of various DAP-DNA complexes (i) DAP-DNA-A, (ii) DAP-DNA-N, (iii) DAP-DNA-H (c) Relative energies of the PDA-DNA complexes in various orientations. As observed in the figure, the stability order is (i) DAP-DNA-A > (ii) DAP-DNA-N > (iii) DAP-DNA-H.; (d) 3D and (e) 2D H-bonded interaction of the DAP with DNA.