卷 71, 编号 4 (2016)

The work describes novel functional bionanosystems for treatment and diagnostics on the basis of proteins, enzymes, polymeric coatings, and magnetic nanoparticles developed at Lomonosov Moscow State University Laboratory for Chemical Design of Bionanomaterials in collaboration with scientists from UNC Eshelman School of Pharmacy (USA). The properties of enzymes (superoxide dismutase, catalase, organophosphate hydrolase, and lysines of bacteriophages) and other drug molecules immobilized in polymeric complexes, as well as the methods for targeted drug delivery using cell-mediated systems and magnetic nanoparticles in in vitro and in vivo operating conditions, are discussed. Physical and chemical characteristics, including data on the functional properties of the nanoformulations, are obtained. The nanoformulations developed demonstrated high potential therapeutic efficacy for the treatment of central nervous system and brain diseases, inflammations (including inflammatory diseases of the eye), cancer and infectious diseases, neurotoxic injury, and others. The possibilities of remote control biochemical reactions using a nonheating low-frequency alternating magnetic field (AMF) for the controlled release of drugs are analyzed in the review. The experimental results of the AMF effects on bionanosystems containing magnetic nanoparticles, such as changing the catalytic activities of enzymes bound to magnetic nanoparticles and ‘disordering’ of the lipid bilayer in membranes, are considered.

The recombinant hybrid protein SR15 composed of streptavidin fused with RGD peptide is obtained. The ability of this protein to recognize human melanoma cells (MeWo line) is demonstrated. Types of expressed RGD-binding integrins in the cells of this line are identified. It is found that the recombinant hybrid protein SR15 binds to integrin αvβ3 on the surface of human melanoma cells. The binding to the protein SR15 results in clustering of αvβ3 receptors on the surface of MeWo cells, internalization of the recombinant protein, and an almost twofold decrease in cell viability.

The oxidative stability of enzymes is mostly dependent on the stability of the Cys and Met residues. Three single point mutants with Met/Leu substitutions in D-amino acid oxidase (DAAO, EC 1.4.3.3) from the yeast Trigonopsis variabilis (TvDAAO) are prepared and characterized. The positions for the amino acid residue substitutions are selected based on multiple alignment of different DAAO amino acid sequences and analysis of the three-dimensional structure of TvDAAO. It is shown that the substrate specificity profile ischanged for all of the mutants. The K_M values for the small and bulky D-amino acidsare increased and decreased, respectively. One of the Met/Leu substitutions results in a two- to threefold increase in thermal stability as compared to the wild-type enzyme. A method for the determination of TvDAAO stability in the presence of hydrogen peroxide is developed and the oxidative stability of wild-type and mutant TvDAAOs is studied. It is shown that none of thethree mutations changes the oxidative stability of the enzymes.

The effect of NO-synthase (NOS) activator and inhibitors on leukemic cell lines HL-60, K-562, and MOLT-4 and bone marrow cells of untreated patients diagnosed with B-cell acute lymphoblastic leukemia compared with lymphocytes from healthy donors is examined. The obtained data on the relationships between the radioprotective, NOS inhibitory, and cytotoxic properties of a number of thiazine, thiazoline, and thiourea derivatives indicates their potential for use as agents for complex radio- and chemotherapy.

Cobalt(II) porphyrazine is synthesized and studied as an active component of a polyvinyl chloride plasticized membrane ion-selective electrodes (ISEs). It is established that regardless of their structure, ISEs are sensitive to iodide. The introduction to the ISE of an ionic additive, ionic liquid 1,3-dihexadecylimidazolium chloride, significantly improves the electrochemical characteristics: the slope of the electrode function reaches −(57 ± 1) mV/dec, c_min = 8.3 × 10^–6 M. Solid-state screen-printed electrodes the surfaces of which are modified by a 1: 4 mixture of cobalt(II) porphyrazine and ionic liquid 1,3-dihexadecylimidazolium chloride demonstrate satisfactory electrochemical characteristics: the slope of the electrode function is −(56 ± 4) mV/dec and c_min = 2.5 × 10^–5 M. The potentiometric selectivity of the ISEs for iodide is studied. It is found that the effect of lipophilic interfering ions is significantly lower for solid state ISEs than for plasticized membrane electrodes.