LOCALIZATION AND ORIENTATION OF METHOXY FLAVONOIDS IN DPPC BILAYERS: EFFECT ON THEIR ANTI-PROLIFERATIVE ACTIVITY
Keywords:Flavones, Flavonols, DPPC, DSC, NMR, Chemical shifts, CSA
Objective: Flavones and flavonols are an important class of naturally occurring flavonoids. They are well known for their pharmacological activity. This activity is associated with the ability of flavones and flavonols to influence membraneâ€“dependent processes. In this paper, we have reported localization, orientation and interaction, of four synthesized flavone/flavonols with 1, 2â€“dipalmitoylâ€“snâ€“glyceroâ€“3â€“phosphocholine (DPPC) bilayers. These are compared with standard flavone; chrysin (CHY) and flavonol Quercetin (QUE).
Methods: The molecules studied are 4Ê¹â€“methoxy flavone (MF), 3Ê¹,4Ê¹â€“dimethoxyflavone (DMF), 4Ê¹â€“methoxyflavonol (MFâ€“ol) and 3Ê¹,4Ê¹â€“dimethoxyflavonol (DMFâ€“ol). The techniques used are Differential Scanning Calorimetry (DSC) and multiâ€“nuclear NMR.
Results: Highest binding to lipid bilayers is shown by DMF, followed by QUE. Based on DSC studies it is seen, that maximum interaction of MF and DMF, takes place with the hydrophobic core of lipid bilayers. DMFâ€“ol shows formation of a heterogeneous system at higher concentrations. The 1H NMR spectra of unilamellar vesicles of DPPC, incorporated with MF, DMF and MFâ€“ol shows significant interaction of these compounds with the alkyl chain of the hydrophobic core. MF, DMF and MFâ€“ol acquire parallel orientation in bilayers with the Bâ€“ring pointing towards hydrophobic core, while DMFâ€“ol acquire mixed orientation. This may be ascribed to the presence of two methoxy and one hydroxyl group on the Bâ€“ring of DMFâ€“ol which hinders its partitioning inside the hydrophobic core of lipid bilayer. Multiâ€“lamellar vesicles (MLV) of DPPC incorporated with flavones, show maximum increase in Chemical Shift Anisotropy in 31P spectrum of DMF. This is followed by MF. DSC.
Conclusion: NMR and binding studies indicate that DMF is partitioned deeply inside the hydrophobic core, while MF, MFâ€“ol and DMFâ€“ol are mostly located in the vicinity of snâ€“glycero region. Therefore, we conclude that DMF which penetrates deepest inside the hydrophobic core also shows the highest antiâ€“proliferative activity against K562 and MCFâ€“7 cancer cell lines. Its activity is also better than CHY.
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