BSA binds the Fe(NO)₂⁺ fragment of DNIC and multiple molecules of [Fe(SC(NH₂)₂)₂(NO)₂]⁺ that prolongs NO donation by this DNIC.

Hydrated copolymers of N-vinylpyrrolidone (VP) with triethylene glycol dimethacrylate as a promising platform for biologically active compounds (BAC) were investigated by different physical chemical methods (dynamic light scattering, infrared spectroscopy, thermal gravimetric analysis, and differential scanning calorimetry) and the quantum chemical modeling of water coordination by the copolymers in a solution. According to the quantum chemical simulation, one to two water molecules can coordinate on one O-atom of the lactam ring of VP units in the copolymer. Besides the usual terminal coordination, the water molecule can form bridges to bind two adjacent C=O groups of the lactam rings of VP units. In addition to the first hydration shell, the formation of a second one is also possible due to the chain addition of water molecules, and its structure depends on a mutual orientation of C=O groups. We showed that N,N-dimethylbiguanidine hydrochloride (metformin) as a frontline drug for the treatment of type 2 diabetes mellitus can be associated in aqueous solutions with free and hydrated C=O groups of the lactam rings of VP units in studied copolymers. Based on the characteristics of the H-bonds, we believe that the level of the copolymer hydration does not affect the behavior and biological activity of this drug, but the binding of metformin with the amphiphilic copolymer will delight in the penetration of a hydrophilic drug across a cell membrane to increase its bioavailability.

Albumin, mucin and oxyhemoglobin act as carriers of the newly synthesized nitrosyl iron complex [Fe₂(SC₆H₃Cl₂)₂(NO)₄] and participate in its transformation.

The cytotoxic activity of a series of dinitrosyl iron complexes (DNICs) with thioureas against cells of different origin has been studied in this work. The cytotoxicity of the studied DNICs proved to be substantially different depending on the structure of the complexes and cell line. Complexes with thiourea and 1,3-dimethylthiourea were found to induce notable cell death in different cell lines of both cancerous and non-cancerous origin, while the N-ethylthiourea-bearing complex induced cell death in cells derived from brain tumors. The studied DNICs effectively release NO while decomposing in solutions, as follows from the electrochemical analysis. It was found that the cytotoxic effects of the studied DNICs did not correlate with their NO-donating ability, hence suggesting that their cytotoxic activity is, in a big part, defined by the long-lived nitrosyl iron–sulfur intermediates formed during the decomposition of the complexes. The structures of the products formed upon hydrolytic decomposition of all studied DNICs have been studied by electrospray ionization mass spectrometry. Stable high-molecular cluster ions containing NO groups namely [Fe₄S₃(NO)₇]⁻ (Roussin's “black salt” anion), [Fe₄S₃(NO)⁵]⁻, [Fe₄S₄(NO)₄]⁻, [Fe₄S₃(NO)₄]⁻ and [Fe₄S₃(NO)₆]⁻ have been detected in the solution of the N-ethylthiourea-bearing complex. The mechanism of Roussin's “black salt” anion formation in a solution of DNIC with N′-ethylthiourea was studied using density functional theory. This moved us near understanding the reasons for the formation of biologically active intermediates upon the decomposition of the complex with N′-ethylthiourea, which are apparently responsible for the unique antiglioma activity of the complex.

The decomposition product of NICs with thiourea and thiosulfate ligands under aerobic conditions can form the high-molecular-weight product Fe(Cys34)(His39)(NO)(NO₂).