Goals and Objectives

• Establishment of a new interdisciplinary laboratory for biohybrid technologies
• Development and application of biohybrids for the targeted delivery of a wide range of drug compounds, including steroids, peptides/proteins, RNA, small organic molecules, and organometallic complexes

Scientific Results

• In the course of the project, new organic molecules capable of inhibiting carbonic anhydrases were synthesized, and a search was conducted for effective blockers of human carbonic anhydrase isoform II activity, which is a validated target for glaucoma therapy. A series of vascular endothelial growth factor (VEGF) inhibitor peptides were synthesized—promising drugs that slow down the formation of the microvascular system in certain eye diseases. Their efficacy was studied using various in vitro and in vivo methods. For effective bioimaging of the drug delivery systems developed in the project, as well as for the future creation of theranostic systems, samples of new phosphorescent labels based on gold, iridium, and platinum complexes were synthesized, carrying reactive groups necessary for covalent conjugation with polymer particles. It was shown that the resulting complexes and their conjugates with polymer carriers possess lifetimes in the microsecond range and the ability to emit in the near-IR region. The resulting compounds were studied in vitro and in vivo, demonstrating their biocompatibility. The synthesis of new polymers—base compounds for the subsequently formed nanoparticles—developed in three main directions: the creation of polymers for small molecule delivery systems, the synthesis and modification of macromolecules to create delivery systems for genetic constructs, and the production of hydrophilic conjugates with anti-VEGF peptides. New polymer colloidal systems suitable for use as drug delivery systems were developed, namely, micro- and nanoparticles based on poly(lactic acid) with a variable number of polyelectrolyte layers of poly(L-lysine) and heparin on their surface; micelles based on amphiphilic chitosan and polyamino acids; and thermo- and pH-sensitive nanogels. Organic linkers were synthesized to function as intermediates between the drug compound and the polymer/nanoparticle/cell surface, as well as polymer chain cross-linking agents designed to ensure controlled cleavage/release of the therapeutic agent upon changes in environmental conditions, such as light radiation or the presence of specific enzymes. All developed materials showed safety for human tissue cells, including retinal and corneal cells, as well as the ability to release the encapsulated drug agent in a controlled manner.
• The possibility of creating biohybrids using two approaches was demonstrated: (a) internalization of nanoparticles loaded with drugs and genetic material into cells; (b) covalent modification of the cell surface with a drug substance using various linkers. According to the first method, poly(lactic acid)-based nanoparticles modified with camelid antibodies were used to encapsulate a new drug for the treatment of multidrug-resistant tuberculosis—perchlozone. The drug dosage received by animals treated with the particles was more than 10 times lower than that of animals receiving the drug orally, which resulted in significantly lower hepatotoxicity of the drug encapsulated in nanocontainers. Conducted studies indicate that the likely mechanism of the targeted action of the drug in particles consists of the uptake of particles by peritoneal macrophages followed by their migration to the site of inflammation via chemotaxis. Methods for controlling particle uptake into cells were developed. According to the second method, a technology for covalent modification of the surface of living T-cells was developed through metabolic labeling with N-azidoacetylmannosamine tetraacetate (Ac4ManNAz) during their in vitro cultivation. Subsequently, a hydrophilic fluorescent dye and selected anti-VEGF peptides modified with dibenzocyclooctyne were bound to the azide groups on the cell surface via an azide-alkyne cycloaddition click reaction. The selected fluorescent dye was used to optimize the biohybrid production technology. Anti-VEGF peptides modified with dibenzocyclooctyne were bound to cells to obtain biohybrid systems capable of detecting and selectively binding the VEGF protein and thus blocking its signaling pathways. It was shown that the modified cells retained their viability.
• Computer algorithms have been developed to model pharmacokinetics following intravitreal injections of polymer materials and biohybrids into the eye. The computer modeling results are in good agreement with pharmacokinetic data obtained through direct measurements of drug concentrations in the vitreous humor. The models account for key parameters such as vitreous volume, the diffusion coefficient of the material in the vitreous, and clearance in the anterior segment of the eye. The models also took into account the polymer degradation rate. Polymer decomposition and the size of the resulting fragments were evaluated in in vitro studies.

Implementation of research results

Organizational and infrastructural transformations

• A new course in English, "Drug Delivery Systems," has been developed and integrated into the educational program for master's students at the SPbU Institute of Chemistry majoring in "Chemistry."
• A new course, "Biomaterials," was developed and included in the educational program for undergraduate students at the SPbU Institute of Chemistry majoring in "Chemistry, Physics, and Mechanics of Materials."
• A laboratory workshop, "Synthesis and Study of Carbon Nanomaterials," was developed and included in the educational program for Master's students at the SPbU Institute of Chemistry majoring in "Fundamental and Applied Aspects of Nanomaterials and Nanotechnologies."
• A laboratory workshop, "Molecular Biology Methods," was developed and tested for Master's students at the SPbU Institute of Chemistry majoring in "Chemistry" (as a supplement to the "Biochemistry" lecture course).

• Meeting of scientific group within Megagrant «Biohybrid technologies for modern medicine», Saint Petersburg, New Peterhof Hotel, December 17-18, 2018.
• «Summer conference on biohybrid technologies», Saint Petersburg, SPbU Graduate School of Management, June 10-11, 2019.

Cooperation

• Department of Chemistry of Natural Compounds, SPbU Institute of Chemistry;
• Department of General and Inorganic Chemistry, SPbU Institute of Chemistry;
• Department of Pathology, Faculty of Medicine, St. Petersburg State University, Saint Petersburg;
• Institute of Experimental Medicine, Saint Petersburg;
• D.O. Ott Research Institute of Obstetrics, Gynecology and Reproductology, Saint Petersburg;
• University of Helsinki, Faculty of Biopharmaceutics, Helsinki, Finland;
• University of Eastern Finland, School of Pharmacy, Kuopio, Finland;
• Leibniz University, Institute of Technical Chemistry, Hannover, Germany;
• Free University, Berlin, Germany;
• University of Florence, Department of Neuroscience, Florence, Italy;
• BIOCAD International Biotechnological Company, Saint Petersburg, Russia.