Liposomal forms of preparations containing biologically active substances extracted from renewable sources of raw materials of plant and animal origin are attracting considerable research and practical interest. Liposomal drugs are efficiently absorbed in the gastrointestinal tract, retaining their structure while passing through the liver and entering the target cell. This allows either smaller amounts of the active substance to be used without losing bioavailability or its effective concentration to be increased.Deer musk liposomes administered at doses equivalent to 300, 1500 and 3000 mg/person/day were found to exhibit an actoprotective effect, whose intensity at the minimum dose (300 mg) is comparable to that achieved at the maximum dose (3000 mg).A course daily transpalatal administration of musk deer liposomes for 14 days was established to statistically significantly improve the indices of physical endurance and working capacity of rats in forced swimming tests under load, kineso-hydrodynamic models of swimming and rotarod performance tests. The observed increase in indicators persisted for another seven days after the cessation of administration, which indicates pharmacological cumulation. The tested drug reduces locomotor and orienting-exploratory activity, at the same time as exhibiting no adverse effect on the psycho-emotional and intoxication status of animals. In addition, no local irritant action on the route of administration was noted. The drug is assigned to toxicity class VI (relatively harmless).

A complex study of the protein composition of the biologically active components of the extracts of the velvet deer antlers (VDA, include Cervus nippon and Cervus elaphus) using two-dimensional gel electrophoresis (2D-PAGE), size-exclusion chromatography (SEC), and peptide mapping with high-performance liquid chromatography with mass spectrometric detection (HPLC-MS) with the use of fermentation by trypsin was done. A molecular-mass distribution characteristic for protein extracts of VDA has been established. Optimal conditions for extraction, chromatographic separation and relative quantitative determination of the main components have been determined. The results of the identification of the most significant (major and minor) protein components in the extracts of the studied objects are described in detail in accordance with the algorithm of the search program Spectrum Mill MS Proteomics Workbench and the protein database Uniprot. The data of protein profiling are clustered according to molecular and biological functions. The connections of the identified proteins with possible mechanisms of biological action and targets, which can be affected by the protein components of the studied objects, are presented. Based on the results of the study, conclusions about the multicomponent protein composition of extracts of VDA were drawn. The marker protein components in the studied extracts are suggested and the possible interrelationships of the detected proteins in the extracts with biological effects are indicated.

In the course of improving the pilot-scale technology for manufacturing a liposomal substance of biologically active substances (BAS) isolated from the gland secretion of musk deer, the authors established operations and modes to ensure the production of liposomes with the specified stability parameters (shape, hydrodynamic diameter, charge, and polydispersity index) in full compliance with GMP requirements and the maximum inclusion of BAS from deer musk into liposomes with the preservation of the initial biological activity of its most important components. The established modes provide for a two-fold reduction in the technological cycle duration with the substance yield of more than 90%. The improved technology ensures the production of at least 80% of liposomes with the main liposome fraction in experimental samples in the range of 200+50 nm and 250+100 nm for the final substance (before lyophilization) and after its recovery (rehydration), respectively, which is considered optimal for transmucosal drug administration. The polydispersity index was less than 0.3, and the zeta (Z)-potential was less than -35 mV. The quality indicators characterizing the dispersity of the resulting liposomal substance were studied by conventional analytical methods (dynamic light scattering, electron microscopy). On the basis of the obtained results, a draft specification for a liposomal substance (powder) containing a complex of BAS isolated from deer musk was finalized.

In order to preserve and increase the biological effectiveness of biologically active substances isolated from deer musk, we studied technological aspects of obtaining a substance of lipid-stabilized stable nanoparticles from deer musk. The stability of the obtained substance was evaluated. It was found that homogenization under high pressure is an optimal approach to obtaining biologically active substances from deer musk. The modes of preparation of a liposomal form of biologically active substances with predetermined dispersion parameters (average particle diameter 250 ± 100 nm, polydispersity index 0.3 ± 0.1, and zeta potential from -5 to -35 mV) were determined. It was found that the high-pressure homogenizer “Donor-5” makes it possible to obtain liposomal dispersions with standard parameters and the degree of inclusion of musk biologically active substances up to 60%, at the same time as providing minimal oxidation and hydrolysis of phospholipids (oxidation index 0.3). Our studies showed that the use of a domestic phosphatidylcholine is economically justified and allows obtaining liposomal forms of proper quality. The quality indicators of the obtained liposomal substance were characterised by conventional analytical methods (dynamic light scattering, electron microscopy, gel chromatography, chromatography-mass spectrometry, etc.). On the basis of the results obtained, a draft specification was developed for a liposomal substance (powder) containing a complex of biologically active substances isolated from deer musk. The developed technology for obtaining a liposomal form of biologically active substances from deer musk can be scaled up in accordance with GMP requirements.