[Show abstract][Hide abstract] ABSTRACT: The clinical application of platinum-based anticancer drugs is greatly limited by severe toxicity. Drug-delivery systems are much sought after to improve the efficacy and applicability of these drugs. Here, we describe a new drug-delivery system comprising a novel platinum complex (bipyridine morpholine dithiocarbamate Pt(II) nitrate) within nanoparticles composed of β-casein (β-CN) and chitosan (CS). The influence of pH on the formation of a colloidally-stable nanocarrier system composed of Pt complex-loaded β-CN and chitosan nanoparticles was investigated using UV-vis spectrometry, dynamic light scattering (DLS) and scanning electron microscopy (SEM). The particles of Pt complex-loaded beta-casein-chitosan formed were stable and soluble in the pH range 5.7-6.2. Hence, the optimal pH for complex formation is between the pI of casein (5.3) and the pKa of chitosan (6.5). DLS data showed that, at both pH values of 5.7 and 6.2, the particles formed had sizes between 200 and 300nm. However, the optimum pH for particle formation was pH 5.7. At this pH, the zeta-potential values of nanoparticles were positive and the particles were stable. SEM analysis confirmed the formation of nanoparticles with good colloidal stability and an average particle size of 200nm. The cytotoxicity of both free and encapsulated Pt complex was evaluated on colorectal carcinoma HCT116 cells. The results obtained indicated that both the cytotoxicity and cellular uptake of platinum were enhanced by its entrapment in β-CN-CS nanovehicles. These findings suggest that this novel drug-delivery system enables drugs to be thermodynamically stable in aqueous solutions and is potentially useful for targeted oral-delivery applications.
[Show abstract][Hide abstract] ABSTRACT: In the present investigation, we have decided to study the interaction between the bovine whey carrier protein of β-lactoglobulin (BLG) with a newly synthesized Pt(II) complex (bipyridine ethyl dithiocarbamate Pt(II) nitrate), as an anti-cancer compound using fluorescence and circular dichroism (CD) spectroscopic methods at two different temperatures of 25 and 37 °C. Also, cytotoxicity and apoptotic activity of this complex have studied against cancer model cell line of K562. Results of intrinsic fluorescence of BLG represent that Pt(II) complex has strong ability to quench the intrinsic fluorescence of protein through dynamic quenching procedure. The values of binding constant (0.21 and 0.27 μM−1 at 25 and 37 °C, respectively) and number of binding site calculated according to the quenching methods at different temperatures. Also, thermodynamic parameters data suggested that hydrophobic interaction plays a major role in the interaction of complex with BLG. In addition, far-UV-CD results show that Pt(II) complex did not induce any significant changes in the secondary structure of BLG. Cytotoxicity and apoptotic effects of the complex toward the cancer cell line of K562 were measured using 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide assay and 4,6-diamidino-2-phenylindole staining methods. From above results, it can be concluded that the bovine whey carrier protein of BLG could bind to be a suitable transfer for this new anti-cancer compound and may be suggested that the anti-tumor activity of this complex reveals typical morphological features of apoptotic death.
Journal of the Iranian Chemical Society 01/2013; 10(5). DOI:10.1007/s13738-013-0232-6 · 1.09 Impact Factor
[Show abstract][Hide abstract] ABSTRACT: A novel monofunctional benzyldithiocarbamate, C(6)H(5)CH(2)NHCSSNa (I), and a bifunctional p-xylidine-bis(dithiocarbamate), NaSSCNHCH(2)C(6)H(4)CH(2)NHCSSNa (II), as sodium salts, were synthesized by reaction between p-xylylenediamine or benzylamine with CS(2) in the presence of NaOH. They were characterized by spectroscopic techniques such as (1)H NMR, IR, and elemental analysis. These water-soluble compounds were examined for their inhibition of both activities of mushroom tyrosinase (MT) from a commercial source of Agricus bisporus. l-3,4- Dihydroxyphenylalanine (L-DOPA) and l-tyrosine were used as natural substrates for the catecholase and cresolase enzyme reactions, respectively. Kinetic studies showed noncompetitive inhibition of I and mixed type inhibition of II on both activities of MT. The inhibition constant (K(I)) of II was smaller than that of I. Raising the temperature from 27 to 37 degrees C caused a decrease in K(I) values of I and an increase in values of II. The binding process for inhibition of I was only entropy driven, which means that the predominant interaction in the active site of the enzyme is hydrophobic; meanwhile, the electrostatic interaction can be important for the inhibition of II due to the enthalpy driven binding process. Fluorescence studies showed a decrease of emission intensity without a shift of emission maximum in the presence of different concentrations of compounds. An extrinsic fluorescence study did not show any considerable change of the tertiary structure of MT. Probably, the conformation of inhibitor-bound MT is stable and inflexible compared with uninhibited MT.
Journal of Enzyme Inhibition and Medicinal Chemistry 09/2009; 25(2):272-81. DOI:10.1080/14756360903179351 · 2.33 Impact Factor
[Show abstract][Hide abstract] ABSTRACT: Three iso-alkyldithiocarbonates (xanthates), as sodium salts, C3H7OCS2Na (I), C4H9OCS2Na (II) and C5H11OCS2Na (III), were synthesized, by the reaction between CS2 with the corresponding iso-alcohol in the presence of NaOH, and examined for inhibition of both cresolase and catecholase activities of mushroom tyrosinase (MT) from a commercial source of Agricus bisporus. 4-[(4-methylbenzo)azo]-1,2-benzendiol (MeBACat) and 4-[(4-methylphenyl)azo]-phenol (MePAPh) were used as synthetic substrates for the enzyme for the catecholase and cresolase reactions, respectively. Lineweaver-Burk plots showed different patterns of mixed and competitive inhibition for the three xanthates and also for cresolase and catecholase activities of MT. For cresolase activity, I and II showed a mixed inhibition pattern but III showed a competitive inhibition pattern. For catecholase activity, I showed mixed inhibition but II and III showed competitive inhibition. These new synthesized compounds are potent inhibitors of MT with K(i) values of 9.8, 7.2 and 6.1 microM for cresolase inhibitory activity, and also 12.9, 21.8 and 42.2 microM for catecholase inhibitory activity for I, II and III, respectively. They showed a greater inhibitory potency towards the cresolase activity of MT. Both substrate and inhibitor can be bound to the enzyme with negative cooperativity between the binding sites (alpha > 1) and this negative cooperativity increases with increasing length of the aliphatic tail in these compounds in both cresolase and catecholase activities. The cresolase inhibition is related to the chelating of the copper ions at the active site by a negative head group (S-) of the anion xanthate, which leads to similar values of K(i) for all three xanthates. Different K(i) values for catecholase inhibition are related to different interactions of the aliphatic chains of I, II and III with hydrophobic pockets in the active site of the enzyme.
Journal of Enzyme Inhibition and Medicinal Chemistry 05/2007; 22(2):239-46. DOI:10.1080/14756360601114536 · 2.33 Impact Factor
[Show abstract][Hide abstract] ABSTRACT: Binding properties and conformational change of human growth hormone (hGH) upon interaction with Fe3+ were investigated at 27 °C in NaCl solution, 50 mM, by calorimetry and spectroscopy. UV spectroscopy indicates that thermal denaturation of hGH is an irreversible process and is accompanied by aggregation. At an optimum concentration of iron thermal denaturation of hGH becomes reversible. Results from equilibrium dialysis and isothermal titration calorimetry indicate a set of four binding sites on hGH for Fe3+. Interaction of three iron ions with hGH prevents irreversibility and aggregation. Differential scanning calorimetry confirms the UV spectroscopic finding. Domain analysis by DSC shows that in the presence of iron, there are at least two main transitions corresponding with the two groups of helices. Deconvolution of the main transitions provides two sub-transitions each, the first pair is similar, but the second pair is considerably different in the enthalpy change of unfolding. Interaction of iron ions with hGH prevents aggregation by an effect on the hydrophobicity and provides information about its structure and thermal denaturation.
[Show abstract][Hide abstract] ABSTRACT: Three new n-alkyl dithiocarbamate compounds, as sodium salts, C4H9NHCS2Na (I), C6H13NHCS2Na (II) and C8H17NHCS2Na (III), were synthesized and examined for inhibition of both cresolase and catecholase activities of mushroom tyrosinase (MT) from a commercial source of Agaricus bisporus in 10 mM phosphate buffer pH 6.8, at 293K using UV spectrophotometry. Caffeic acid and p-coumaric acid were used as natural substrates for the enzyme for the catecholase and cresolase reactions, respectively. Lineweaver-Burk plots showed different patterns of mixed and competitive inhibition for catecholase and cresolase reactions, respectively. These new synthetic compounds can be classified as potent inhibitors of MT due to Ki values of 0.8, 1.0 and 1.8 microM for cresolase inhibitory activity, and also 9.4, 14.5 and 28.1 microM for catecholase inhibitory activity for I, II and III, respectively. They showed a greater potency in the inhibitory effect towards the cresolase activity of MT. Both substrate and inhibitor can be bound to the enzyme with negative cooperativity between the binding sites (alpha > 1) and this negative cooperativity increases with increasing length of the aliphatic tail in these compounds. The inhibition mechanism is presumably related to the chelating of the binuclear coppers at the active site and the different Ki values may be related to different interaction of the aliphatic chains of I, II and III with the hydrophobic pocket in the active site of the enzyme.
Journal of Enzyme Inhibition and Medicinal Chemistry 08/2005; 20(4):393-9. DOI:10.1080/14756360500179903 · 2.33 Impact Factor